SPARK 2024 Abstracts

Ava Teng

Cedars-Sinai

Mentor: Dr. Maedeh Mozneb, PhD

PI: Dr. Arun Sharma, PhD

Poster #001

Poster #001

Title

Effect of β-Estradiol on GCaMP-hiPSC-Cardiomyocyte Viability

Abstract

Cardiovascular diseases remain a leading cause of morbidity and mortality worldwide. Human-induced pluripotent stem cells (hiPSCs) offer promising opportunities to develop new therapies in the cardiac regenerative medicine field. However, many cardiac stem cell research pursuits exclude sex-specific variability, and the limited research on these differences exacerbates gender biases in healthcare. Thus, we examine the cardiotoxic effect of β-estradiol, the most potent form of estrogen found in humans, on GCaMP-hiPSC-cardiomyocytes (GCaMP-hiPSC-CMs), a genetically modified reporter cell line capable of intrinsically showing calcium signaling. We hypothesized that β-estradiol would enhance cardiomyocyte electrophysiology and health. Our preliminary findings suggest that β-estradiol has a protective effect, evidenced by lowered cytotoxicity indicated by lower lactate dehydrogenase (LDH) release levels and higher CCK8 cell metabolic readout compared to the control. However, variability in calcium signaling frequency and amplitude points to complex β-estradiol effects on signal transduction pathways. Further research is needed to elucidate these mechanisms.

Colin Hsiang-Hsien Hwang

Cedars-Sinai

Mentor: Vladimir Zhemkov, PhD

PI: Clive Svendsen, PhD

Poster #002

Poster #002

Title

MS2-MBP Pull-down Assay to Investigate G4C2 RNA Interaction in Cells

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disease characterized by degeneration of motor neurons. A genetic link has been shown between abnormal intronic G4C2 repeat expansion (n>20) in C9ORF72 gene and a genetic form of ALS, but the exact mechanisms behind this are unclear. Earlier, we discovered that G4C2 RNA repeats colocalize with U12 minor spliceosome snRNA. We use an MS2-tagging approach to investigate how G4C2 repeats interact with cellular RNAs. Constructs containing G4C2-n25 repeats fused to an MS2-tag, or control constructs (MS2-tag alone) were cloned and expressed in mammalian cells. MS2-MBP protein was expressed in bacteria, purified, and attached to amylose beads. RNAs bound to MS2-tagged transcripts were captured using an MS2-MBP pull-down assay. Interacting RNAs were analyzed by RT-PCR. More minor than major spliceosome snRNAs were bound to the G4C2-MS2 relative to the MS2 control, suggesting G4C2 repeats interact directly with minor spliceosome snRNAs in cells.

Daniel Svediani

Cedars-Sinai

Mentor: Ruchi Shah, Ph.D.

PI: Alexander V. Ljubmiov, Ph.D.

Poster #003

Poster #003

Title

Epigenetic therapies improve diabetic corneal wound healing and stem cells

Abstract

Diabetes mellitus (DM) appears to be an epigenetic disease, which causes epithelial alterations including impaired wound healing due to dysfunctional limbal epithelial stem cells (LESC) in the cornea. Previously, we used a nanobioconjugate (NBC) and DNA demethylator zebularine to modulate DM-altered protein expression such as cathepsin F (increased in DM), c-Met, and Wnt-5a (decreased in DM), to stimulate wound healing and LESC marker expression. Here, we used combined therapy with the NBC and zebularine. In human diabetic organ-cultured corneas (n=6) wounded with 1-heptanol- we performed western blot and immunostaining. These corneas treated with the drug combination suppressed the expression of cathepsin F and increased c-Met and Wnt-5a expression, resulting in significantly improved wound healing and increased expression of LESC markers (K15, K17, ABCG2), diabetic markers (nidogen-1 and integrin α3β1), and wound healing mediators (p-Akt and p-p38). Therefore, combined epigenetic therapy may have potential in treating diabetic corneal epithelial wound healing.

Ethan Sung

Cedars-Sinai

Mentor: Edward Novinbahkt

PI: Celine Riera, PhD.

Poster #004

Poster #004

Title

Propranolol Reduces Brown Adipose Tissue Thermogenesis in Response to Cold

Abstract

Obesity is one of the leading metabolic diseases prevalent in today’s society and has the highest comorbidity rate with many other life-threatening conditions. The presence and activation of brown adipose tissue (BAT) in both humans and mouse models has proven to be effective in combating obesity through the increase of energy expenditure in response to a variety of pharmacological and environmental cold stimuli. This study aims to understand how cold signals are processed by the central nervous system and how these signals are integrated to regulate metabolism and cold-induced thermogenesis. We discovered that a common beta-blocker, propranolol, is effective in reducing the thermogenic capacity of BAT in response to cold in mice, either directly through adipose tissue or indirectly via neural circuits in the brain. Future experiments manipulating adrenergic receptors using gene therapy can hold the keys for resistance against obesity and other forms of metabolic disease.

Irene Thomas

Cedars-Sinai

Mentor: Paul Linesch, PhD

PI: Joshua Breunig, PhD

Poster #005

Poster #005

Title

Efficacy of Various Genomic Safe Harbor Sites on Transgene Expression

Abstract

Genomic safe harbor (GSH) sites are locations in a cell's genome where a transgene can be stably integrated, without disturbing the expression of other genes. In gene therapy, these sites can be utilized to integrate engineered donor DNA to treat medical conditions with a genetic basis, such as cancer and other genetic conditions. This project investigates the expression of the GreenLantern (GL) fluorescent protein reporter transgene in different GSHs within the genome of mouse Neural Stem Cells (mNSCs) and human stem cells (hiPSCs). The GSH sites in the hiPSCs are in the AAVS1 and ROGI loci, while the mNSCs site is in the ROSA26 locus. These 3 strains were cultured, and the expression of the GL transgene was analyzed using fluorescence and qPCR. By investigating which GSH site results in greater expression levels, we can improve the effectiveness of gene therapies that rely on transgene integration.

Keith Mahar

Cedars-Sinai

Mentor: Namdev More Ph.D.

PI: Dmitriy Sheyn Ph.D.

Poster #006

Poster #006

Title

Microgel Cell Therapy for Intervertebral Disc Degeneration Treatment

Abstract

Intervertebral disc degeneration is the breakdown of the discs cushioning the spine. Current invasive treatments provide only temporary relief carrying significant risk, highlighting the need for cell therapy to restore disc function. Nucleus pulposus cell (NPC) therapy aids intervertebral disc degeneration treatment by replenishing the cellular matrix. Challenges with NPC therapy include cell survival, integration, and degradation before therapy. This study evaluates the influence of microgel size on cell viability and extracellular matrix production. NP cells were encapsulated in FibMA microgels using 100µm and 200µm channel widths, producing 150µm and 400µm microgels, respectively. Encapsulation efficiency and cell viability were equally suitable across sizes, with marker expressions for COL1, ACAN, and SOX9 remaining stable by day 14. Microgels stored at 37°C showed a superior cell viability compared to 4°C and 25°C, suggesting favorable storage conditions. These findings support the potential of FibMA microgels in enhancing NPC therapy efficacy through size optimization.

Madison Cheungsomboune

Cedars-Sinai

Mentor: Emily A. Hatanaka

PI: Joshua J. Breunig, PhD

Poster #007

Poster #007

Title

Characterization of Immune Populations in an Adult GBM Mouse Model

Abstract

Glioblastoma is the most common and aggressive brain tumor in adults. Despite aggressive treatment of surgical resection, radiation therapy (RT) and temozolomide chemotherapy, patient prognosis is 14 months. The Breunig Lab models brain tumors in mice utilizing MADR technology. In this study, we modeled adult glioblastoma aiming to investigate the immune microenvironment. We performed immunohistochemistry on RT treated/untreated tumor-bearing mice to identify presence of peripheral and resident macrophages. Additionally, peripheral immune niches such as the femur and skull bone marrow were assessed to classify immune populations using flow cytometry. We observed at 7 days post-RT, decrease in tumor size in response to RT and higher concentration of macrophages in the tumor. In distal bone marrow niches, we observed a higher proportion of Ly6G high granulocytes in the skull compared to femur in tumor-bearing mice. These findings help to better characterize immune niches in glioblastoma and understand immune responses to RT.

Sarah Martellino

Cedars-Sinai

Mentor: Michael Ramos

PI: Ritchie Ho, PhD

Poster #008

Poster #008

Title

Cell-Specific Transgene Expression in Spinal Cord of SOD1-G93A Mice

Abstract

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease where motor neurons (MNs) degenerate. The SOD1-G93A mouse is an animal model of hereditary ALS containing over 20 copies of the mutant human (hSOD1) gene randomly inserted into the mouse genome, causing MN degeneration and paralysis by 160 days of age. Previously, we found upregulated SOD1 mRNA during ALS progression from single-nuclei transcriptomes. However, it was unclear whether these were endogenous mice (mSod1) or transgenic hSOD1 transcripts. Here, we investigated hSOD1 mRNA and protein expression in SOD1-G93A mouse spinal cords during ALS progression. Using qPCR, we observed hSOD1 but not mSod1 mRNA is upregulated. Unexpectedly, hSOD1 protein aggregates were only present in the spinal cord ventral area MNs at the presymptomatic age, but were not present in the end-stage. Furthermore, diffuse hSOD1 protein is increased in end-stage astrocytes. These findings suggest complex transcriptional and translational processes regulating hSOD1 expression during ALS.

Selin Bilgi

Cedars-Sinai

Mentor: Dr. Antonio C Fuentes-Fayos; PhD

PI: Dr. Joshua J Breunig; PhD

Poster #009

Poster #009

Title

Understanding the Activation of Transcription Factor Etv4 in K27M Gliomas

Abstract

Pediatric brain tumors, particularly K27M pediatric high-grade glioma (HGG), are among the most aggressive and challenging cancers in children. Here, we used a mouse model known as MADR, which precisely mimics the clinical human phenotype, together with functional genomics, to study the role of the activation of transcription factor, Etv4. To do that, in vivo/in vitro approaches were used, using Doxycycline as an expression inducer. Our results showed that in vivo induction was insufficient to observe the role of Etv4. Thus, we established organoids from non-induced mice. After the validation and induction in vitro, we observed that Etv4 expression in vitro increased the number of organoids versus tagBFP control organoids under No Doxycycline/Doxycycline conditions. The induction efficacy of Dox was determined through fluorescent microscopy and flow cytometry. To conclude, merging MADR with functional genomics is useful to unveil the role of Etv4 as a potential oncogene in K27M pediatric glioma.

Urja Parekh

Cedars-Sinai

Mentor: Brijesh Singh, PhD

PI: Ritchie Ho, PhD

Poster #010

Poster #010

Title

Inhibition of EZH1/2 promotes the maturation of iPSC-derived motor neurons

Abstract

Amyotrophic lateral sclerosis is characterized by a gradual and debilitating deterioration of motor neurons as patients age. Despite the revolutionary impact of induced pluripotent stem cell (iPSC)-based disease modeling on medical research, newly differentiated motor neurons are inherently too immature to exhibit ALS disease phenotypes such as proteinopathies and cell death, undermining their effectiveness as models of molecular pathology. Previous research suggests that slow maturation in early phases of iPSC-derived motor neuron differentiation may be attributed to epigenetic suppression of genes that promote neuronal development and maturation. To alleviate this obstacle, we used a small molecule inhibitor of EZH1 and EZH2, the catalytic subunits of Polycomb repressive complex 2 that tri-methylate histone 3 at lysine 27, which in turn mediate gene silencing. EZH1/2 inhibition resulted in enhanced neuronal maturation, supporting the potential of epigenetic modifiers in iPSC-derived neurons for higher fidelity modeling of neurodegenerative diseases. 

Samaria Bailey

Charles R. Drew University of Medicine and Science

Mentor: Pranabananda Dutta, PhD

PI: Jaydutt Vadgama, PhD

Poster #011

Poster #011

Title

Investigating Cancer Stem Cell Biomarkers in Triple-Negative Breast Cancer

Abstract

This study investigates the expression of cancer stem cell markers ALDH1A1, CD44, OCT4, and Nanog in triple-negative breast cancer (TNBC), an aggressive subtype lacking estrogen, progesterone, and HER2 receptors. Using chemiluminescent Western blotting on TNBC cell lines and analyzing The Cancer Genome Atlas breast cancer datasets, we aimed to understand these markers' contribution to TNBC's aggressive behavior and their potential as therapeutic targets. Results showed strong ALDH1A1 and CD44 expression in all TNBC cell lines, with ALDH1A1 particularly high in the Basal subtype. OCT4 and Nanog were undetected. ALDH1A1 levels were remarkably high in MDA-MB-468 and HCC1937 cells, and moderate in MDA-MB-231 cells. These findings suggest ALDH1A1 and CD44 as promising targets for TNBC treatment, while OCT4 and Nanog may be less relevant. This research enhances our understanding of TNBC molecular characteristics and guides new treatment approaches.

Brianna Boye

Charles R. Drew University of Medicine and Science

Mentor: Eiji Yoshihara, PhD and Jinhyuk Choi, PhD

PI: Eiji Yoshihara, PhD

Poster #012

Poster #012

Title

Human Pancreatic β-cell Proliferation

Abstract

Diabetes is a metabolic disease that is characterized by uncontrolled blood sugar levels, damaging blood vessels. Insulin is responsible for controlling the pathway of glucose, but controlling the mass of insulin-producing cells, β-cells, during diabetes progression is unpredictable. However, the regulatory mechanisms of β-cell proliferation is poorly understood. This research aims to provide a solution for those suffering from diabetes through regenerative medicine. In diabetic patients, pancreatic β-cells no longer function. A method proposed by Yoshihara Lab is to generate healthy β-cells from hPSCs. The data collected from molecule screening of hPSC-derived β-like cells identified the novel small molecules, which stimulates β-cell proliferation. The lab confirmed its effect in clonal human β-cell line, EndoC-βH1. EndoC-βH1 mimics functioning β-cells in the pancreas, with a homogeneity of 99%. This research emphasizes the importance of regenerative mechanisms to proliferate and regenerate pancreatic β-cells, which can help scientists develop better strategies to combat diabetes.

Emma Do

Charles R. Drew University of Medicine and Science

Mentor: Pranabananda Dutta, PhD

PI: Jaydutt Vadgama, PhD

Poster #013

Poster #013

Title

Expression of CSC Biomarkers in Luminal A/B and HER2-Enriched Breast Cancer

Abstract

Breast cancer (BC) is a heterogeneous disease with subtypes such as luminal A, luminal B, and HER2-positive. Cancer stem cell (CSC) markers help identify CSCs that play a crucial role in cancer initiation, growth, and recurrence. The research aimed to explore the expression of CSC markers, such as OCT4, Nanog, CD44, and ALDH1A1, in BC subtypes and tumor-derived cell lines. Chemiluminescent Western blot with cell lysate prepared from cell lines representing the subtypes and probing with primary antibody. Analyze The Cancer Genome Atlas (TCGA) breast cancer datasets on cBioPortal for CSC marker mRNA expression. The data suggests presence of the CD44 marker in MCF-7, JIMT-1, HCC1954, SKBR3, and AU565 cell lines. ALDH1A1, OCT4, and Nanog protein were undetected. In the TCGA dataset, luminal A had the highest mRNA expression for CD44 and Nanog. The CD44 marker is highly expressed in some luminal A, luminal B, and HER2-positive BC subtypes.

Kenneth Garnett

Charles R. Drew University of Medicine and Science

Mentor: Thomas Lozito, PhD

PI: Thomas Lozito, PhD

Poster #014

Poster #014

Title

Assessing the Ability of Lizards to Regenerate Digit Tips

Abstract

Lizards have the natural ability to regenerate their tails amputation, but the extent to which lizards can regrow their digit tips is largely unknown. Two drugs, Smoothened Agonist (SAG) and cyclopamine, have been known to significantly affect the formation of blastemas for lizard tails. SAG is known to induce the regeneration of lizards’ tails with more cartilage than normal, whereas cyclopamine induces blastemas with no cartilage. The aim of this research is to assess whether or not lizards are able to regenerate digit tips after the introduction of cyclopamine and SAG. The study involved 15 adult green anoles (Anolis carolinensis) treated with either cyclopamine, SAG, or vehicle control. The results of the study were pending at the time of abstract submission. However, based on initial observations, it seems that the lizard's fingertips will not experience any regeneration for either of the two groups.

Brian Higgins

Charles R. Drew University of Medicine and Science

Mentors: Yong Wu, MD, PhD and Ke Wu, MD, PhD

PI: Jaydutt Vadgama, PhD

Poster #015

Poster #015

Title

MSC’s Influence on the Proliferation and Invasion of TNBC Cells

Abstract

Recent studies suggest that mesenchymal stem cells (MSCs) may influence cancer progression, but their specific role in Triple-negative breast cancer (TNBC remains unclear. This project aims to explore how MSCs impact the proliferation and invasion of TNBC cells and to investigate the underlying mechanisms that may alter their behavior, potentially identifying new therapeutic targets. MSCs were co-cultured with MDA-MB-468 cells to assess changes in cell proliferation and invasion using MTT assays, wound healing, and transwell invasion assays. Additionally, molecular analyses were conducted to identify signaling pathways. Preliminary data indicate that MSCs significantly enhance both proliferation and invasion capabilities of TNBC cells. Molecular analyses suggest involvement of multiple signaling pathways in mediating these effects. These insights provide a foundation for identifying novel therapeutic targets aimed at mitigating MSC-mediated progression of TNBC.

Anaiya Mayfield

Charles R. Drew University of Medicine and Science

Mentors: Yong Wu, MD, PhD and Ke Wu, MD, PhD

PI: Jaydutt Vadgama, PhD

Poster #017

Poster #017

Title

Mesenchymal Stem Cells Induce Chemoresistance in TNBC Cells

Abstract

Triple-negative breast cancer (TNBC) is characterized by its aggressive behavior and limited treatment options, posing significant clinical challenges. Emerging evidence suggests that mesenchymal stem cells (MSCs) may play a role in inducing chemoresistance in cancer cells. This study aims to investigate the potential of MSCs in promoting chemoresistance in TNBC, specifically by examining their impact on the sensitivity of TNBC cells (MDA-MB-468) to the chemotherapy drug SN38. MSCs will be co-cultured with MDA-MB-468 TNBC cells and treated with SN38. Changes in cell viability and apoptosis will be assessed using MTT assays and flow cytometry. Molecular analyses will be conducted to elucidate the underlying signaling pathways involved in MSC-induced chemoresistance. Preliminary data suggest that MSCs induce chemoresistance in MDA-MB-468 cells against SN38, resulting in reduced apoptosis and enhanced cell viability. Our study highlights the crucial role of mesenchymal stem cells (MSCs) in fostering chemoresistance in triple-negative breast cancer (TNBC) cells.

Nicolas Mendoza

Charles R. Drew University of Medicine and Science

Mentor: Shehla Pervin, PhD

PI: Shehla Pervin, PhD

Poster #018

Poster #018

Title

Increased Resistance of a Subset of MCSCs to NO-Mediated Cell Death

Abstract

Breast cancer is a complex disease with multiple subtypes, often involving mammary cancer stem cells (MCSCs) that contribute to tumor initiation and progression. MCSCs, marked by ALDH1 and CD44, are heterogeneous, plastic, and resistant to chemotherapy, making them key targets for treatment. Macrophages are innate immune cells that are present in M1 phenotype that is anti-tumor or M2 phenotype that promotes tumor growth. This research investigated whether M1 macrophages can reduce MCSC populations in the triple-negative breast cancer cell line HCC 1806. RAW 264.7, a mouse macrophage line, was stimulated to express inducible nitric oxide synthase (iNOS). HCC 1806 cells, grown in specific conditions to promote MCSC formation as mammospheres, showed reduced ALDH1 expression when co-cultured with stimulated macrophages. However, a subset of cells resisted nitric oxide-induced death. The tudy concludes that nitric oxide alone is insufficient to eliminate MCSCs, suggesting the need for combination therapies.

Estephanie Salinas

Charles R. Drew University of Medicine and Science

Mentor: Thomas Lozito, PhD

PI: Thomas Lozito, PhD

Poster #019

Poster #019

Title

Hedgehog Signaling in Lizard Tail and Mouse Digit Tip Blastemas

Abstract

Hedgehog Signaling pathway (Hh pathway) is involved in regulating cell proliferation and differentiation during lizard tail regrowth. Previous studies demonstrate that treating amputated lizard tails with the Smoothened Agonist (SAG) activates Hh signaling during tail regrowth. The purpose of this research is to determine whether Hh signaling plays similar roles during mouse digit tip regeneration, the other main model of amniote appendage regeneration. To conduct this study, regenerated tail samples were collected from green anole lizards (Anolis carolinensis), and regenerated digit tip samples were collected from mice (Mus musculus). Animals were treated with either SAG, Cyclopamine, or vehicle control every 48 hours, and samples were collected 14 days post-amputation. Lizard tail and the mouse digit tip samples were analyzed by histology/in situ hybridization for Hh signaling markers. Results are pending at time of abstract submission, but we hypothesize that regenerating lizard tails, not mouse blastemas, respond to Hh signaling.

EwongoAbasi Umoh

Charles R. Drew University of Medicine and Science

Mentor: Shehla Pervin, PhD

PI: Shehla Pervin, PhD

Poster #020

Poster #020

Title

Cross-talk between Mammary Cancer Stem Cells and Differentiated Adipocytes

Abstract

Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer that is responsible for the majority of breast cancer deaths, of which African-American women are disproportionately affected. African-American TNBC tumors have a higher concentration of cancer stem cells; stem cells have a high potency for self-renewal, tumor initiation, and progression. Furthermore, the progression of breast cancer is influenced by adipocytes, and higher concentrations of differentiated adipocytes are directly adjacent to TNBC tumors. This study demonstrates an interaction exists between 1806 mammary cancer stem cells and differentiated adipocytes, which leads to aggressive tumor behavior. Co-cultured differentiated adipocytes with 1806 mammary cancer stem cells; performed a western blot. There was an upregulation of Sox 2 & ALDH1A1 that indicates cancer stem cell renewal. Further, the high expression of pERK & pAKT indicates the activation of pathways that promote cancer cell proliferation, survival, and metastasis.

Charlie Perez

Children’s Hospital Los Angeles

Mentors: G. Esteban Fernandez PhD, Michael Schumacher PhD

PI: G. Esteban Fernandez PhD, Michael Schumacher PhD

Poster #021

Poster #021

Title

THE EPITHELIAL RESPONSE OF INFLAMED INTESTINAL TISSUE IN MOUSE MODEL IBD

Abstract

Globally, almost 5 million individuals suffer from inflammatory bowel disease (IBD). Only 40% of patients report symptom relief with current treatments. We sought to determine whether the cellular composition of the intestinal epithelium changes during IBD to understand how that may contribute to disease progression. We stained colon tissue from mice with anti-IL-10R induced colitis and 2 and 4 weeks after recovery to identify epithelial cells, dividing cells, tuft cells, secretory cells, and goblet cells. We observed consistent staining only for proliferating cells and quantified their number in colonic crypts. We observed increased cell proliferation in mice recovering from colitis, indicating that changes in the composition of the intestinal epithelium may be occurring, but the identity of cell population(s) in flux remains unclear. In the future, the staining protocol needs to be optimized for the other cell type markers and to stain small intestine tissue.

Bianca Chirrick

Children’s Hospital Los Angeles

Mentor: Vini John, PhD

PI: Mark R. Frey, PhD

Poster #022

Poster #022

Title

The role of neuregulin growth factors in intestinal regeneration aft

Abstract

Inflammatory Bowel Disease (IBD) is characterized by chronic ulceration in the GI tract, frequently in the colon. We hypothesized that the proteins NRG1 and NRG2 promote wound healing of the colonic epithelial barrier. To test this, we made experimental wounds in cultured colonic epithelial cells, stimulated with EGF (positive control), NRG1, or NRG2, photographed them over time, and measured percent closure. Then we used qPCR to detect possible restitution markers in wounded and unwounded cultures. Interestingly, both NRG1 and NRG2 enhanced wound healing as compared to control and EGF. Moreover, RNA coding for the healing/tight junction protein Cldn4 was increased in cells stimulated by NRG2; in contrast, NRG1 stimulation appeared to increase Muc2 expression, a common marker for goblet cells. Our data suggest that neuregulins induce both gene expression changes and cellular behavior necessary for repair and regeneration. These findings could be an avenue for developing future IBD treatments.

Bernice Nkemnji

Children’s Hospital Los Angeles

Mentors: G. Esteban Fernandez PhD, and Michael Schumacher PhD

PI: G. Esteban Fernandez PhD, and Michael Schumacher PhD

Poster #023

Poster #023

Title

THE EPITHELIAL RESPONSE OF INFLAMED INTESTINAL TISSUE IN MOUSE MODEL IBD

Abstract

Globally, almost 5 million individuals suffer from inflammatory bowel disease (IBD). Only 40% of patients report symptom relief with current treatments. We sought to determine whether the cellular composition of the intestinal epithelium changes during IBD to understand how that may contribute to disease progression. We stained colon tissue from mice with anti-IL-10R induced colitis and 2 and 4 weeks after recovery to identify epithelial cells, dividing cells, tuft cells, secretory cells, and goblet cells. We observed consistent staining only for proliferating cells and quantified their number in colonic crypts. We observed increased cell proliferation in mice recovering from colitis, indicating that changes in the composition of the intestinal epithelium may be occurring, but the identity of cell population(s) in flux remains unclear. In the future, the staining protocol needs to be optimized for the other cell type markers and to stain small intestine tissue.

Isabella Martinez

Children’s Hospital Los Angeles

Mentors: Velia Alcala, Osanna Kosoyan, and Valentina Villani, PhD

PI: Laura Perin, PhD

Poster #024

Poster #024

Title

Smartflares: Illuminating Pathways in Wilms Tumor Research

Abstract

Wilms Tumor (WT) is a pediatric kidney cancer that develops due to overexpression of SIX2. SIX2 is a transcription factor that’s important for nephron formation in development. In physiological conditions, cells expressing SIX2 are exhausted before birth. In WT, SIX2 activity continues past birth, causing cells to rapidly replicate and form tumors. To investigate SIX2+ cells, we used a tool called Smartflares which detect RNA targets in live cells without damaging the cells. They’re composed of a gold nanoparticle, capture strand, and reporter strand. We hypothesized that we could isolate SIX2+ cancer stem cells from cultured WT cells. We labeled cultured WT cell populations with Smartflares targeting SIX2 and performed FACS analysis, finding >60% of the cells were SIX2+. Confirmation of SIX2 protein expression was through Western Blotting. Successful isolation of these cells will allow us to study the mechanisms that regulate Wilms Tumor.

Allison Paredes

Children’s Hospital Los Angeles

Mentor: Vini John, PhD

PI: Mark R. Frey, PhD

Poster #025

Poster #025

Title

The role of neuregulin growth factors in intestinal regeneration after inju

Abstract

Inflammatory Bowel Disease (IBD) is characterized by chronic ulceration in the GI tract, frequently in the colon. We hypothesized that the proteins NRG1 and NRG2 promote wound healing of the colonic epithelial barrier. To test this, we made experimental wounds in cultured colonic epithelial cells, stimulated with EGF (positive control), NRG1, or NRG2, photographed them over time, and measured percent closure. Then we used qPCR to detect possible restitution markers in wounded and unwounded cultures. Interestingly, both NRG1 and NRG2 enhanced wound healing as compared to control and EGF. Moreover, RNA coding for the healing/tight junction protein Cldn4 was increased in cells stimulated by NRG2; in contrast, NRG1 stimulation appeared to increase Muc2 expression, a common marker for goblet cells. Our data suggest that neuregulins induce both gene expression changes and cellular behavior necessary for repair and regeneration. These findings could be an avenue for developing future IBD treatments.

Jennifer Ramirez Flores

Children’s Hospital Los Angeles

Mentors: G. Esteban Fernandez PhD, Michael Schumacher PhD

PI: G. Esteban Fernandez PhD, Michael Schumacher PhD

Poster #026

Poster #026

Title

THE EPITHELIAL RESPONSE OF INFLAMED INTESTINAL TISSUE IN MOUSE MODEL IBD

Abstract

Globally, almost 5 million individuals suffer from inflammatory bowel disease (IBD). Only 40% of patients report symptom relief with current treatments. We sought to determine whether the cellular composition of the intestinal epithelium changes during IBD to understand how that may contribute to disease progression. We stained colon tissue from mice with anti-IL-10R induced colitis and 2 and 4 weeks after recovery to identify epithelial cells, dividing cells, tuft cells, secretory cells, and goblet cells. We observed consistent staining only for proliferating cells and quantified their number in colonic crypts. We observed increased cell proliferation in mice recovering from colitis, indicating that changes in the composition of the intestinal epithelium may be occurring, but the identity of cell population(s) in flux remains unclear. In the future, the staining protocol needs to be optimized for the other cell type markers and to stain small intestine tissue.

Serena Hannifin

Children’s Hospital Los Angeles

Mentors: Velia Alcala, Osanna Kosoyan, Valentina Villani, PhD

PI: Laura Perin, PhD

Poster #027

Poster #027

Title

Smartflares: Illuminating Pathways in Wilms Tumor Research

Abstract

Wilms Tumor (WT) is a pediatric kidney cancer that develops due to overexpression of SIX2. SIX2 is a transcription factor that’s important for nephron formation in development. In physiological conditions, cells expressing SIX2 are exhausted before birth. In WT, SIX2 activity continues past birth, causing cells to rapidly replicate and form tumors. To investigate SIX2+ cells, we used a tool called Smartflares which detect RNA targets in live cells without damaging the cells. They’re composed of a gold nanoparticle, capture strand, and reporter strand. We hypothesized that we could isolate SIX2+ cancer stem cells from cultured WT cells. We labeled cultured WT cell populations with Smartflares targeting SIX2 and performed FACS analysis, finding >60% of the cells were SIX2+. Confirmation of SIX2 protein expression was through Western Blotting. Successful isolation of these cells will allow us to study the mechanisms that regulate Wilms Tumor.

Isabella Krauss

Children’s Hospital Los Angeles

Mentors: Hripsime Chomoyan, MD, and Hasmik Soloyan, MD

PI: Sargis Sedrakyan, PhD

Poster #029

Poster #029

Title

Stem Cell Extracellular Vesicles Protect Glomerular Endothelial Cell

Abstract

Alport Syndrome (AS) is a form of chronic kidney disease caused by mutations in genes encoding type IV collagens. Metabolic dysfunction of the glomerular endothelial cells (GEC) contributes to AS progression. Here, we describe how vascular endothelial growth factor-A (VEGF-A) affects mitochondrial biogenesis and the potential of using amniotic fluid stem cell (AFSC) derived extracellular vesicles (EVs) to restore GEC homeostasis. Primary cultures of human GEC were exposed to different concentrations of VEGF and were treated with AFSC-EVs. Mitochondrial biogenesis markers were evaluated by Western blot and immunohistochemistry at 24, 48, and 72 hours. At 24 hours post VEGF-A exposure, the mitochondrial biogenesis marker PGC-1⍺ was decreased in GEC, which was reversed upon treatment with EVs. Thus, we have demonstrated the potential of AFSC-EVs to rescue mitochondrial biogenesis in GEC which may lead to the development of targeted new therapies for AS and other forms of CKD.

Kaitlyn Morrison

Children’s Hospital Los Angeles

Mentor: Mohammadreza Kasravi, MD/MBA

PI: Stefano Da Sacco, PhD

Poster #030

Poster #030

Title

Optimizing Podocyte Differentiation of Amniotic Fluid Progenitor Cells

Abstract

odocytes—cells essentially for kidney function—are irreversibly damaged in Alport syndrome (AS). We have been testing whether amniotic fluid stem cells (AFSC) can be induced to form podocyte-like cells as a possible replacement therapy. Based on prior work showing that hypoxia induces the podocyte marker WT1 in Wilms Tumor cells, we tested whether the hypoxia mimetic cobalt chloride induces podocyte markers in cultured AFSC. We treated AFSC in vitro with cobalt chloride and are performing western blot and immunohistochemical analysis for WT1, HIF-1⍺, and nephrin. If successful, these experiments will identify a potential cell based therapy approach for AS.

Anna Villatoro

Children’s Hospital Los Angeles

Mentor: Mohammadreza Kasravi, MD/MBA

PI: Stefano Da Sacco, PhD

Poster #031

Poster #031

Title

Optimizing Podocyte Differentiation of Amniotic Fluid Progenitor Cells

Abstract

Podocytes—cells essentially for kidney function—are irreversibly damaged in Alport syndrome (AS). We have been testing whether amniotic fluid stem cells (AFSC) can be induced to form podocyte-like cells as a possible replacement therapy. Based on prior work showing that hypoxia induces the podocyte marker WT1 in Wilms Tumor cells, we tested whether the hypoxia mimetic cobalt chloride induces podocyte markers in cultured AFSC. We treated AFSC in vitro with cobalt chloride and are performing western blot and immunohistochemical analysis for WT1, HIF-1⍺, and nephrin. If successful, these experiments will identify a potential cell based therapy approach for AS.

Seth Pickens

Children’s Hospital Los Angeles

Mentors: G. Esteban Fernandez PhD, Michael Schumacher PhD

PI: G. Esteban Fernandez PhD, Michael Schumacher PhD

Poster #032

Poster #032

Title

THE EPITHELIAL RESPONSE OF INFLAMED INTESTINAL TISSUE IN MOUSE MODEL IBD

Abstract

Globally, almost 5 million individuals suffer from inflammatory bowel disease (IBD). Only 40% of patients report symptom relief with current treatments. We sought to determine whether the cellular composition of the intestinal epithelium changes during IBD to understand how that may contribute to disease progression. We stained colon tissue from mice with anti-IL-10R induced colitis and 2 and 4 weeks after recovery to identify epithelial cells, dividing cells, tuft cells, secretory cells, and goblet cells. We observed consistent staining only for proliferating cells and quantified their number in colonic crypts. We observed increased cell proliferation in mice recovering from colitis, indicating that changes in the composition of the intestinal epithelium may be occurring, but the identity of cell population(s) in flux remains unclear. In the future, the staining protocol needs to be optimized for the other cell type markers and to stain small intestine tissue.

Obianuju Oyeoka

Children’s Hospital Los Angeles

Mentors: Hripsime Chomoyan, MD, Hasmik Soloyan, MD

PI: Sargis Sedrakyan, PhD

Poster #034

Poster #034

Title

Stem Cell Extracellular Vesicles Protect Glomerular Endothelial Cell

Abstract

Alport Syndrome (AS) is a form of chronic kidney disease caused by mutations in genes encoding type IV collagens. Metabolic dysfunction of the glomerular endothelial cells (GEC) contributes to AS progression. Here, we describe how vascular endothelial growth factor-A (VEGF-A) affects mitochondrial biogenesis and the potential of using amniotic fluid stem cell (AFSC) derived extracellular vesicles (EVs) to restore GEC homeostasis. Primary cultures of human GEC were exposed to different concentrations of VEGF and were treated with AFSC-EVs. Mitochondrial biogenesis markers were evaluated by Western blot and immunohistochemistry at 24, 48, and 72 hours. At 24 hours post VEGF-A exposure, the mitochondrial biogenesis marker PGC-1⍺ was decreased in GEC, which was reversed upon treatment with EVs. Thus, we have demonstrated the potential of AFSC-EVs to rescue mitochondrial biogenesis in GEC which may lead to the development of targeted new therapies for AS and other forms of CKD.

Kristy Xiao

UCSF Benioff Children’s Hospital Oakland

Mentors: Mohammad H. Mehraban PhD and Aras N. Mattis, MD PhD

PI: Aras N. Mattis, MD PhD

Poster #035

Poster #035

Title

Knocking out miR-432 in Induced Pluripotent Stem Cells (iPSCs)

Abstract

Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) affects ~25% of people worldwide. MASLD is characterized by abnormal hepatic lipid accumulation. This abnormality induces endoplasmic reticulum stress, triggering inflammation, and potentially progressing to Metabolic Dysfunction-Associated Steatohepatitis (MASH). The gene TM6SF2 is crucial in regulating liver lipid metabolism. Recently it was proved that miR-432 regulates TM6SF2 by degrading TM6SF2 mRNA. Our study investigates the effects of miR-432 knockout on TM6SF2 expression and lipid accumulation in iPSCs. MiR-432 in wildtype iPSCs was knocked out with CRISPR-Cas9 and confirmed through Sanger sequencing and ICE analysis. RT-qPCR studies showed that the depletion of miR-432 in iPSCs results in increased TM6SF2 expression. When challenged with oleic acid and stained with Nile red, iPSCs with miR-432 knockout displayed reduced lipid accumulation, suggesting miR-432 knockout increases TM6SF2 mRNA stability. Understanding TM6SF2 mRNA stability's impact provides insights into MASLD and MASH pathogenesis and potential therapeutic approaches targeting liver lipid metabolism.

Nicole Padilla Alvarado

UCSF Benioff Children’s Hospital Oakland

Mentor: Meagan Esbin, PhD

PI: Robert Tijan, PhD

Poster #036

Poster #036

Title

Trying to see where and when MFSD2A and Syncytin 2 bin

Abstract

The growth of the placenta requires a special cell that needs to fuse with other cells that form a syncytium. To form the syncytium there are two important cell membrane proteins called MFSD2A and Syncytin 2. Syncytin2 initiates fusion while MFSD2A is a receptor. While we know that MFSD2A and Syncytin 2 bind together during cell-cell fusion we don’t know the timing and location when the two proteins bind together. We want to create fluorescently tagged MFSD2A constructs by using bacterial cloning. Then we will test if combining the cells with cell-cell fusion in live human cells will work using our engineered MFSD2A. The next step would be to see where and how MFSD2A and Syncytin 2 bind together. The significance is that we want to learn how cell-cell fusion works in order to understand why placental diseases happen.

Nghi Nham

UCSF Benioff Children’s Hospital Oakland

Mentors: Shahrbanoo Keshavarz, PhD and Aras N. Mattis, MD PhD

PI: Aras N. Mattis, MD PhD

Poster #037

Poster #037

Title

iPSC phenotypes after reversion of SUGP1 rs10401969 SNP

Abstract

Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) is a liver condition that is initiated by excess hepatic lipid accumulation affecting over 30% of adults worldwide. MASLD can advance to Metabolic Dysfunction-Associated Steatohepatitis (MASH), fibrosis, and cirrhosis. Previous research demonstrated strong linkage between SNPs within the chromosome and excess lipid accumulation within hepatocytes- characterizing the SUGP1 gene as a regulator of cholesterol metabolism. In recent in vitro studies, iPSC derived Hepatocytes with SUGP1 knockdown significantly increased lipid accumulation intracellularly. Thus, we aim to analyze phenotypes within iPSC patient lines after reversion of SUGP1 rs10401969 SNP through in vitro analysis of MASH-carrier, non-MASH control(s), and SUGP1 clone corrected SNP iPSC lines challenged with Oleate. Intracellular phenotypes of corrected lines are expected to decrease in lipid accumulation comparable to healthy iPSC lines contrary to MASH-carrier iPSC lines. This research will further establish if SUGP1 is a therapeutic target for patients with MASLD.

Belem Osorio

UCSF Benioff Children’s Hospital Oakland

Mentors: Yuqing Zhang PhD and Marisa Medina PhD

PI: Marisa Medina, PhD

Poster #038

Poster #038

Title

Differentiating Induced Pluripotent Stem Cells (iPSCs) into β-cells

Abstract

Statins are a class of cholesterol lowering drugs used to prevent cardiovascular disease, however, some statins users develop new-onset type 2 diabetes (NOD). While diabetes can be caused by insufficient insulin secreted from pancreatic β-cells, it is unclear whether statin-induced NOD is due to impaired insulin secretion. The Medina Lab has created iPSCs from statin-induced NOD cases vs. controls and has implemented a differentiation protocol to produce pancreatic β-like cells (iPSC-β). We first sought to determine whether iPSCs from NOD cases vs. controls can be differentiated with the same efficiency. We created iPSC-β from a NOD case and control donor and tested for expression of β-cell markers using flow cytometry. While the iPSC from the NOD case had higher differentiation efficiency compared to the control, this finding cannot be generalized due to inadequate sample size. Additional studies are ongoing to validate the iPSC-β model for future studies on NOD.

Mercy Niyi-Awolesi

UCSF Benioff Children’s Hospital Oakland

Mentors: Jenny Lee; Ke Li, PhD; Brian Shy, MD, PhD

PI: Brian Shy, MD PhD

Poster #039

Poster #039

Title

Optimizing a Non-Viral, Knock-in Approach in Hematopoietic Stem Cells

Abstract

Objective: Identify optimal conditions for clinically compatible, non-viral knock-in in HSC populations by optimizing 3 components: safe harbor targets, pulse codes, and HDRT concentration. Results: Between the safe harbor targets, the AAVS1 site showed higher knock-in rates than GS94. An increase in pulse code energy levels (HSC 3 to 5) also led to an increase in knock-in rates and toxicity, leading to a comparable number of cells with knock-in in all three pulse codes tested. Additionally, increasing the HDRT concentration also increased our knock-in rates as expected. However, our highest amount of HDRT (2 pmol) seemed to lead to higher levels of toxicity when also using higher pulse code energy levels. Further optimization efforts are needed to increase knock-in rates.

Vy Phan

UCSF Benioff Children’s Hospital Oakland

Mentors: Yuanyuan Qin, PhD and Marisa Medina, PhD

PI: Marisa Medina, PhD

Poster #040

Poster #040

Title

Validating iPSC Differentiation to iPSC-Heps Using Stage-Specific Markers

Abstract

Induced pluripotent stem cells (iPSCs) are pluripotent cells that can be differentiated to create iPSC-derived hepatocyte-like cells (iPSC-Heps), an important tool for modeling disease risk. Prior to the study, iPSC-Heps were often authenticated to have hepatocyte-like characteristics by confirming their expression of ALB and ASGPR, two proteins that are specifically expressed in hepatocytes. The goal of this study was to determine whether commonly used commercially available antibodies against ALB and ASGPR demonstrated specificity for hepatocyte-like cells by comparing their expression levels in undifferentiated iPSCs (which should have no detectable levels of these proteins) with HepG2, a human hepatoma cell line that expresses both proteins. We tested two ASGPR antibodies and three ALB antibodies and found that only one of each showed the expected positive detection in HepG2 and negative detection in iPSC. These findings demonstrate the importance of authenticating molecular markers used during iPSC-Hep validation.

Katherine Bien

City of Hope

Mentor: Chelsea Jang

PI: Yingfeng Deng, Ph.D.

Poster #041

Poster #041

Title

Evaluating the impact of Xbp1s on beta cell survival

Abstract

In individuals with Type 2 Diabetes (T2D), sustained high insulin production induces endoplasmic reticulum stress which results in beta cell loss. Studies have shown that such beta cell loss may be reversible. However, the mechanism for reversing this loss remains unclear. This study aims to assess if beta cell loss due to Xbp1s, an ER stress transducer is reversible. Overexpression of Xbp1s is induced in BIX mice by a doxycycline diet. After 5 weeks of the doxycycline diet feeding, half of the mice are switched to a regular diet for 5 weeks to recover. Beta cell mass is quantified using Immunofluorescent staining of insulin in mouse pancreata. Beta cell mass is expected to decrease in mice on the doxycycline diet but recovered in mice on the regular diet. Our study will reveal how Xbp1s modulates beta cell survival, which can help in identifying novel therapeutic targets to prevent T2D. 

Isaac Jung

City of Hope

Mentor: Margarita Gutova, M.D., Ph.D.

PI: Russell Rockne, Ph.D.

Poster #042

Poster #042

Title

Impact of Extracellular Vesicles on CAR-T therapy for Glioblastoma

Abstract

Glioblastoma is a highly aggressive form of brain tumor. Conventional forms of cancer therapy are ineffective due to the blood-brain barrier, a layer of blood vessels and tissue that protects the brain from harmful substances. Chimeric Antigen Receptor T-cells(CAR-T), genetically modified T-cells that target specific tumor antigens, represent a promising strategy to treat Glioblastoma uninhibited by the blood-brain barrier. Studies show that Glioblastoma's tumor microenvironment(TME) hampers CAR-T therapy. Extracellular Vesicles(EVs), are lipid-bound vesicles secreted by cells to transport biomolecules and can encourage immunosuppression. However, EV’s effect on the TME and CAR-T efficacy is poorly understood. We conducted an in-vitro study to research these effects, collecting data about glioblastoma growth with varying CAR-T and EV concentrations. I aim to model EV’s impact on CAR-T-Glioblastoma interactions through differential equations and system dynamics analysis. Conclusions will clarify how CAR-T cells interact with EVs, allowing improvement of CAR-T therapy against Glioblastoma.

Jonathan Li

City of Hope

Mentor: Balint Beres

PI: Tijana Jovanovic-Talisman, Ph.D

Poster #043

Poster #043

Title

Deep learning-based methods for characterization of extracellular vesicles

Abstract

Extracellular vesicles (EVs) are nanoscale membrane structures shed from cells that contain proteins, nucleic acids, and metabolites. Recent studies have shown that EVs are critical in physiological and pathological processes and intercellular communication. Additionally, EVs represent promising biomarker candidates, as they report rapidly on cell health and can be sampled from accessible biofluids. However, the heterogeneity of EV subpopulations and the small fraction of EVs in patient biofluids originating from diseased cells makes EV subpopulation characterization difficult. To address some of these challenges, this study combines affinity isolation of EVs with two super-resolution imaging methods: Single Molecule Localization Microscopy (qSMLM) and enhanced Super-Resolution Radial Fluctuation (eSRRF). We used deep learning-based processing methods to advance the identification and segmentation of single EVs imaged with super-resolution microscopy methods. Moreover, we used regression models trained on qSMLM and eSRRF colocalization data to benchmark eSRRF’s capability for the multi-parametric characterization of single EVs.

Christine Lin

City of Hope

Mentor: Tammy Chang, M.S.

PI: Jiing-Kuan Yee, Ph.D.

Poster #044

Poster #044

Title

Generating safe and hypoimmunogenic beta-like cells for diabetes treatment

Abstract

Type 1 diabetes (T1D) results from the patient’s immune system attacking insulin-producing β cells in their pancreatic islets, impairing glucose metabolism. Prior studies discovered that differentiating pluripotent stem cells (PSCs) into islet-like structures, called SC-islets, can compensate for lost β cells when transplanted into T1D patients. However, two obstacles remain with this approach: teratoma formation caused by undifferentiated or partially differentiated PSCs and host immunity against SC-islets. Tumor formation is minimized by the conditional expression of a suicide gene such as gene-encoding herpes thymidine kinase (TK). Immunity against SC-islets is addressed through chronic use of immunosuppressants, increasing risks of infection and malignancy in patients. To avoid using immunosuppressors, we will utilize CRISPR-Cas9 gene editing to prevent HLA class I expression from cell surfaces, which we hypothesize will reduce host rejection of SC-islets. Results from this study can enhance SC-islet transplantation in T1D treatments.

Jessica Lu

City of Hope

Mentor: Anthony Park, Ph.D.

PI: Saul Priceman, Ph.D.

Poster #045

Poster #045

Title

Impact of Bead to T Cell Ratio on CAR T Cell Function

Abstract

Chimeric antigen receptor (CAR) T cell therapy involves the engineering of a receptor that selectively targets antigens on the surface of cancerous cells to treat patients with solid tumors and blood cancers. During the manufacturing process of CAR T cells, CD3/CD28 dynabeads are used to activate and expand T cells prior to lentivirally transducing them to express a CAR. While the standard bead: T cell ratio used in cell therapy is 3:1, these CAR T cells exhibit non-target based killing against certain tumor cell lines. Thus, we hypothesized that off-target killing by T cells expanded with a 3:1 bead: tumor ratio could be a result of overactivation. We aimed to lower CAR-T cell overactivation and prevent off-target activity by reducing bead:T cell ratios during CAR-T cell production. Results from this study may improve CAR T cell specificity for better therapeutic efficacy and safety in cancer patients.

Daniel Nam

City of Hope

Mentor: Sneha Varghese, Ph.D.

PI: Sangeeta Dhawan, Ph.D.

Poster #046

Poster #046

Title

DNA damage vulnerability of beta- and alpha-cells in a model of T2D

Abstract

Blood glucose is regulated by insulin-producing beta-cells and glucagon-producing alpha-cells, located in pancreatic islets. Type 2 Diabetes (T2D), often associated with obesity, stems from failure of beta-cells to meet the body's insulin needs. T2D-associated metabolic stress is known to exacerbate DNA damage in beta-cells, but it remains unclear if alpha-cells are similarly affected. Previous research from the Dhawan lab shows that although beta/alpha-cells in developing islets are vulnerable to DNA damage, alpha-cells are more prone due to higher replication and inefficient DNA repair. T2D-associated metabolic stress is known to exacerbate DNA damage. Thus, this study aims to compare DNA damage vulnerability between beta/alpha-cells in T2D. Using antibodies for DNA damage, replication, beta-cells, and alpha-cells, the immuno-stained pancreatic sections from db/db (T2D model) and db/+ (control) mice will be quantified for DNA damage, replication, and damage in replicating beta/alpha-cells, providing insights into differential DNA damage susceptibility of islets in T2D.

Natalie Sandoval

City of Hope

Mentor: Heather N. Zook, Ph.D.

PI: Teresa Ku, Ph.D.

Poster #047

Poster #047

Title

Investigation of 3 Compounds on Human Ductal Suspension Cell Proliferation

Abstract

The pancreas has three main types of cells: acinar, ductal, and islet cells. In type 1 diabetes (T1D), the immune system destroys islet beta cells, which produce insulin in response to high blood sugar. To restore the ability to produce insulin, we aim to utilize progenitor-like cells from human pancreatic ductal cells, which can self-renew and differentiate into functional beta-like cells. To increase ductal progenitor-like cell numbers, we conducted high-throughput screening of >3000 FDA-approved compounds. Of these, three lead compounds increased Adenosine tri-phosphate (ATP), a cellular energy source. Changes in ATP levels can result from altered cell numbers or metabolic state, and I hypothesize that the lead compounds increased cell numbers by proliferation. I will test my hypothesis using immunofluorescence staining and quantification of proliferation markers in treated and untreated cells. My results will aid future efforts to use ductal progenitor-like cells as beta cell replacement therapy for T1D.

Jordan Struve

City of Hope

Mentor: Malaney Young, Ph.D.

PI: Robert J. Hickey, Ph.D.

Poster #048

Poster #048

Title

Overcoming Cisplatin Resistance in Ovarian Cancer with AOH1996

Abstract

Ovarian cancer patients who undergo chemotherapy with cisplatin often develop resistance to treatment, resulting in cancer recurrence. Our lab has discovered a compound, AOH1996, which selectively kills cancer cells via targeting a cancer-associated isoform of proliferating cellular nuclear antigen (caPCNA). Preliminary data shows that AOH1996 can overcome cisplatin resistance in ovarian cancer cells. We have developed a series of cisplatin-resistant ovarian cancer cell lines to investigate the ability of AOH1996 to overcome their acquired resistance using sulforhodamine B assay. We will explore the mechanism of acquired resistance and AOH sensitivity by quantifying the relative expression levels of caPCNA and c-myc in these cells through 2D gels and Western Blot analysis. This project will enhance our understanding of AOH1996’s therapeutic application and how caPCNA and c-myc expression levels are implicated in overcoming the resistance of ovarian cancer cells to cisplatin.

Julier Thein

City of Hope

Mentor: Cesar Gonzalez

PI: Qiang Lu, Ph.D.

Poster #049

Poster #049

Title

Elucidating the role of Zeb1 during the embryonic gliogenic switch

Abstract

Orderly production of the major types of neural cells, including neurons and glial cells, is required to give rise to the functional central nervous system and promote homeostasis in the brain. During early brain development, neural progenitor cells (NPCs) follow an intrinsic biological clock to produce neurons and glial cells in tandem. How the transition from neurogenesis to gliogenesis is regulated during brain development is not fully understood. We previously identified a set of transcription factors through single-cell RNA sequence analysis that may regulate the switch between this transition. Of the factors found, Zeb1 emerged as a promising candidate. To better understand and characterize the role of Zeb1 during the gliogenic switch, we will perform in vitro Zeb1 gain and loss of function studies using murine and human NPCs. These results will contribute basic knowledge of glial cells and may lead to therapies for glial cell-based diseases.

Angelina Wu

City of Hope

Mentor: Babgen Manookian, Ph.D.

PI: Andrei Rodin, Ph.D.

Poster #050

Poster #050

Title

Spatial Gene Expression in Glioblastoma via Information-Theory Methods

Abstract

With recent advancements in gene sequencing, ST provides quantitative genetic information with a spatial component. AI techniques, like machine learning and deep learning, effectively identify patterns from large datasets, but limited explainability and interpretability of these (often ‘black-box’) models calls for new methods to dissect and delineate cellular mechanisms. We created an interpretable information-theoretic computational method to study spatial genetic expression and cellular dynamics in brain glioblastoma. Using benchmark models, we developed a baseline algorithm to analyze spatial entropy distribution with a kernel that scans across tissue samples. This method is applied to two mouse brain glioblastoma ST datasets from the NIH Gene Expression Omnibus database, providing insights into tumor microenvironments and glioblastoma stem cell dynamics. The primary deliverable is a novel generalizable and interpretable methodology for differentiating between homogeneous and spatially organized gene expressions to advance our understanding of cell function, fate, and differentiation.

Aaron Yang

City of Hope

Mentor: Leying Zhang, Ph.D.

PI: Behnam Badie, M.D.

Poster #051

Poster #051

Title

Enhancing Glioblastoma Treatment Through RAGE Inhibitor Variants

Abstract

Glioblastoma, a class of cancer that causes formation of malignant brain tumors, impedes conventional cancer therapies due to the brain’s unique microenvironment. The receptor for advanced glycation end products (RAGE) is a transmembrane receptor known to regulate glioma inflammatory response. Prior research has demonstrated that through knockout of the RAGE gene, certain discovered drugs can inhibit the NF- κB signaling pathway, preventing cell damage by mitigating inflammation. However, subsequent tests show that these drug therapies impair adaptive immune response due to inadvertent downregulation of interferon regulatory factors (IRFs). Thus, we aim to identify drug variants that precisely target anti-inflammatory response while preserving the IRF pathway. Our study evaluates chemically modified RAGE-inhibitors in vitro within THP1 cells, which permit simultaneous monitoring of the NF-κB and IRF pathways, to assess RAGE expression and quantity of RAGE ligands. By highlighting effective RAGE-inhibitors, results from this study will enhance anti-tumor efficacy of glioblastoma treatment.

Alenabell Yang

City of Hope

Mentor: Hongmin Wu, Ph.D.

PI: Zuoming Sun, Ph.D.

Poster #052

Poster #052

Title

Evaluating the Effect of RORyt mediated Th17 cells in T1D

Abstract

Type 1 Diabetes (T1D) is an autoimmune disease where a patient’s immune cells–leukocytes responsible for activating the immune response–attack insulin-producing pancreatic β-cells. While Th1 cells are known to be a contributor to T1D, the function of Th17 cells in disease onset remains controversial. Using mouse models, this study investigates the therapeutic potential for targeting retinoic acid receptor-related orphan receptor gamma t (RORγt), the key regulator of Th17 cells, in T1D. Nonobese diabetic mice were fed a diet containing the RORγt inhibitor IMU-935 then monitored. Mice administered IMU-935 showed lower levels of Th17 cells in the pancreatic lymph node and reduced incidence of diabetes and insulitis. To understand the direct effect of RORγt mediated Th17 cells in T1D, we will perform plasma autoantibody analysis and mRNA analysis on the extracted spleens and pancreata. Enhancing knowledge on the impact of RORγt inhibitors in T1D holds promise for therapeutic treatments against the disease.

Vivian Yuan

City of Hope

Mentor: Ignacio Norambuena-Soto, Ph.D.

PI: Zhao Wang, Ph.D.

Poster #053

Poster #053

Title

Diabetic cardiomyopathy in vitro model using iPSC-derived cardiomyocytes

Abstract

Diabetic cardiomyopathy (DCM) is a leading cause of mortality in individuals with diabetes. The high glucose level adversely affects the structure and function of the heart and causes cardiac dysfunction. While animal models are widely used to investigate the underlying molecular mechanism, limitations exist in their ability to mimic human-specific aspects of DCM. The use of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) represents a promising approach for developing patient-specific models. However, its application in DCM remains unexplored. Here, we aim to establish the iPSC-CM culture model for the future use in diabetes-related cardiovascular research. To mimic DCM in vitro, we will treat iPSC-CMs with high glucose and palmitate. Then, we will analyze pathological changes including insulin resistance, metabolic shift, and cell death. Developing a human DCM in vitro model will greatly accelerate drug discovery, illuminate new signaling pathways, and improve clinical outcomes for patients with diabetes.

Kira Zeng

City of Hope

Mentor: Wilma Tixi, Ph.D.

PI: Hung-Ping Shih, Ph.D.

Poster #054

Poster #054

Title

The Role of SREBF2 in Pancreatic Islet Development

Abstract

Human pluripotent stem cells (hPSCs) can be differentiated into pancreatic islet cells and transplanted into patients to treat type 1 diabetes (T1D). However, the challenge is getting the hPSC-derived islet cells to function. Our studies demonstrated that islet function require the cell adhesion protein Ctnna1. Inactivation of Ctnna1 negatively impacts islet aggregation and function. We also found that Ctnna1 activity promotes cholesterol biosynthesis (CBS). What remains unclear is the role that CBS plays in islet aggregation and function. To address this, we investigated the requirement of Sterol Regulatory Element Binding Transcription Factor 2 (Srebf2), a master regulator promoting CBS, in developing islets. We genetically deleted Srebf2 in developing mouse islets and studied how the islet aggregation and structures may be affected. Our study could provide insights into the role of Srebf2 during islet development and inform the new strategies to improve hPSC differentiation for T1D therapies.

Dyllan Fernandez

UC San Diego

Mentor: Benjamin Gastfriend, PhD

PI: Richard Daneman, PhD

Poster #055

Poster #055

Title

ZIC (Zinc finger of the cerebellum) and EBF (Early B-cell Factor) interacti

Abstrac

tMural cells are specialized cells that help maintain strength and integrity for blood vessels while also modulating blood flow within the central nervous system. ZIC1 and ZIC4 transcription factors are specifically expressed in and may be responsible for the development of central nervous system (CNS)mural cells. Along with ZIC1 and ZIC4, Early B-cell factor (EBF1) is expressed in CNS mural cells despite also being expressed in non-CNS mural cells. We sought to understand the interactions between ZIC and EBF1 transcription factors in CNS mural cells. To test, we expressed tagged versions of the ZIC and EBF1 proteins and used co-immunoprecipitation and western blotting to determine whether they interacted. Here, we demonstrated that ZIC4 has interactions with EBF1, while ZIC1 does not. These findings may stimulate further research in ZIC and EBF1 transcription factors, particularly in mice.

Joannah Jugalbot

UC San Diego

Mentors: Hung-Chi Tu, Ph.D, Po-Yuan Chen, Ph.D and Jose Chacon, B.S

PI: David Traver, Ph.D

Poster #056

Poster #056

Title

PhD Investigating Notch Ligands’ Necessity in Hematopoietic Stem Cell Forma

Abstract

Hematopoietic stem cells (HSCs) are vertebrate cells that can self-renewal and generate all blood cells. During HSCs development, endothelial cells acquire hematopoietic potential to become HSCs by signaling cues from the niche environment. Cell-cell interactions such as Notch signaling are integral for cell fate decisions, cell differentiation, and the emergence of HSCs. Understanding the hematopoietic precursors’ interaction with their environment reveals the complexities of notch signaling’s role in hematopoiesis. Our study dissects the role of notch ligands Dlb and Jagged1b and their effect on HSC development using zebrafish as our model organism. We conducted Dlb and Jagged1b knockdown experiments via morpholino injections on Gata2b: kalta4; UAS: LA-eGFP transgenic zebrafish, where HSCs are marked in GFP. Thereupon, annotation of cell counts in knockdown and control conditions are executed to elucidate the necessity of Dlb and Jagged1b. The results will advance our understanding of the molecular environment supporting HSC development.

Kimberly Vincent

UC San Diego

Mentor: Phuong Le, PhD

PI: Eugene Yeo, Ph.D

Poster #057

Poster #057

Title

Identifying Changes in Synaptic Ribosome Proteins in Amyotrophic Lateral Sc

Abstract

Amyotrophic lateral sclerosis (ALS) is a rare neurodegenerative disease characterized by muscle weakness and gradual degradation of motor neurons in the central nervous system, which control muscles responsible for voluntary movements. As ALS progresses, these muscles begin to weaken and atrophy, ultimately leading to death by respiratory failure. Previous studies have demonstrated that the Kinesin Family Member 5A (KIF5A) protein is a motor neuron-binding protein that binds to mRNA-encoding synaptic ribosomes. Because of this, mutations in the KIF5A protein could potentially contribute to synaptic changes, leading to ALS in neurons. My project involves examining how synaptic numbers change in healthy compared to ALS iPSC-derived motor neurons by utilizing stem cell techniques and immunofluorescence. By comparing the differences of synaptic ribosome proteins in neurons from healthy individuals compared to those with ALS, we aim to better understand these differences. With this, we can develop potential therapeutics for this devastating disease.

Victor Martinez

UC San Diego

Mentor: James Bartram PhD

PI: Robert Signer PhD

Poster #058

Poster #058

Title

R and Bioinformatics: A Pipeline for Data Analysis

Abstract

Leukemia is a common blood cancer in the United States and poses significant challenges for patients and families. Overall, leukemia takes the lives of one in three patients diagnosed. Despite efforts, the research done does not yet promise a cure or reversal of leukemia. In our lab, we use R to rapidly manage and convert data such as genotypes into usable information leading to hundreds of genetic samples analyzed simultaneously, accelerating research exponentially. We have built a script that can quickly analyze differences in immunologic response between mice with genetic differences in K60R and STI, two genes that play a role in tumor suppression. Overall, we have made a pipeline to optimize efficiency, which can be adapted to fit other datasets. This optimization will benefit this testing phase and many more to come. This useful pipeline will accelerate the foundational processes required for the future analysis that comes with research.

Brook Zerihun

UC San Diego

Mentors: Inglis Chandra, Huang Hsu-Ching, and Janae A Hardy

PI: Dr. Sandeep Patel

Poster #059

Poster #059

Title

EGFR Co-Mutation by Ethnicity in Non-Small-Cell Lung Cancer

Abstract

Lung cancer is one of the leading causes of cancer mortality globally. Non-small-cell lung cancer (NSCLC) is one of the most common types, with adenocarcinoma being the most prevalent, accounting for over 70 percent of all NSCLC diagnoses. Squamous cell carcinoma and large cell carcinoma account for the remainder percentage of NSCLC. One of the most notable mutations in NSCLC is an epidermal growth factor receptor (EGFR) mutation. It is known that EGFR, one of the first identified biomarkers in lung cancer, drives abnormal cell growth. Tyrosine kinase inhibitors also known as TKI drugs have been developed against various specific EGFR mutations; it is unknown what this means in our diverse patient base. We aim to better understand co-mutation burden in patients with NSCLC EGFR mutations by comparing diverse demographics, including the patient's race and/or ethnicity.

REFERENCE LINKS

The co-mutation of EGFR and tumor-related genes leads to a worse prognosis and a higher level of tumor mutational burden in Chinese non-small cell lung cancer patients - PMC (nih.gov)

Jamie Ramos

UC San Diego

Mentor: Rhea Robertson, PhD

PI: Kathleen Fisch, PhD

Poster #060

Poster #060

Title

Modeling Genetic Causes of Preeclampsia with Trophoblast Stem Cells

Abstract

The placenta is responsible for gas/nutrient exchange between the mother and the baby. Preeclampsia (PE) is obstetric high blood pressure that occurs in 3-8% of pregnancies and can result from abnormal placental development, restricting fetal growth. The placenta is made of trophoblast stem cells (TSCs) that differentiate into extravillous trophoblasts (EVTs) and syncytiotrophoblasts (STBs). The causes of PE are not well understood; however, we hypothesize that mutations in certain genes may affect the ability of trophoblast cells to differentiate and proliferate, causing placental dysfunction underlying PE. To test our hypothesis, we will use CRISPR-Cas9 gene editing technology to introduce mutations in iPC-derived TSCs in previously identified genes of interest. We will then perform assays to characterize the function of mutated TSCs, STBs, and EVTs. The long-term goal is to identify molecular mechanisms underlying placental dysfunction in PE and uncover preventative measures and treatment options.

Jonathan Methippara

UC Riverside

Mentors: Iris Andrade, Ph.D. Student and Simon Andal, Undergraduate Researcher

PI: Sonali Chaturvedi, Ph.D.

Poster #061

Poster #061

Title

The Autorepression Circuit in HSV-1 Dictates the Fate of the Virus

Abstract

Herpes simplex virus-1 (HSV-1) is the number one cause of infectious blindness. Encephalopathy associated with HSV-1 poses a grave challenge for neonates, as 10/13 of infants infected with HSV-1 show signs of significant brain lesions. Patients with HSV-1 undergoing organ transplantation procedures have a higher likelihood of rejection, jeopardizing their lives. HSV-1 codes for an autorepression circuit by forming a negative feedback loop on its indispensable ICP4 protein. The disruption of this autorepression circuit reduces viral fitness by “open-loop lethality”. The applications of disrupting the autorepression circuit can be used therapeutically (Chaturvedi et al. Cell 2022). We amplify the IE gene of Cytomegalovirus and optimize conditions to amplify the ICP4 gene of HSV-1. We will create a stable cell line containing the ICP4 autorepression circuit using lentivirus, followed by testing the autorepression circuits in constantly evolving environments to understand how these circuits dictate the fate of the virus.

Brian Chen

Stanford

Mentor: Jinyi Xiang MD/PhD

PI: Irving Weissman MD

Poster #068

Poster #068

Title

Investigating Long-Term Hematopoietic Stem Cell Dynamics in 3D Culture

Abstract

Long term hematopoietic stem cells (LT-HSCs) are essential for the continuous renewal of the blood and immune systems. Residing in their bone marrow niche, LT-HSCs require various factors to sustain their populations and support their fate decisions to meet system demands. To study these niche factors in vitro, we established 3D bone marrow culture systems from Hoxb5-tdTomato LT-HSC reporter mice, allowing live tracking of Hoxb5+ LT-HSCs. Using this in vitro method, we observed the long-term survival of most bone marrow cell types. Notably, LT-HSCs were maintained throughout the observation period, predominantly as single cells within the 3D cell culture systems, closely mimicking their natural bone marrow microenvironment. Additionally, some LT-HSC divisions were observed in the cultured bone marrow systems. This method provides an excellent in vitro model to study the fundamental biology of LT-HSCs and the niche factors that control their fate.

Alberto Alonzo Garcia Brito

Stanford

Mentor: Yuan Guan PhD, and Wenlong Rong PhD

PI: Gary Peltz MD/PhD

Poster #069

Poster #069

Title

BRD4 Inhibitor's Effect on Liver Epigenomics

Abstract

Liver fibrosis is a life-threatening disease that results from the excessive accumulation of extracellular matrix proteins, such as collagen, which disrupts the normal structure and function of the liver. It results from chronic liver injury due to heavy alcohol or fat consumption or viral infection. The liver wound-healing response activates hepatic stellate cells to transform into myofibroblast-like cells that produce collagen and as fibrosis progresses, the liver's ability to perform essential functions is compromised leading to more complications for the patient. There are currently no approved treatments that can reverse liver fibrosis. We used human hepatic organoids (HOs) generated from induced Pluripotent Stem Cells (iPSC) that were treated with TGF-b, as a model system for liver fibrosis. We used these HOs to examine the effect of BRDR4 inhibitors on developing liver fibrosis. Our results demonstrate that BRD4 inhibitors, like JQ1, could be effective therapies for the treatment of liver fibrosis.

Saleh Saleh

Stanford

Mentor: Suyash Raj BA

PI: Irving Weissman MD

Poster #070

Poster #070

Title

Modeling Trichloroethylene Induced Neurotoxicity

Abstract

Trichloroethylene (TCE) is a colorless, non-flammable, volatile liquid commonly found in chemical cleaners, adhesives, and degreasers. While it was recently banned by the FDA, many patients who have been exposed and will continue to deal with damage resulting from TCE exposure. This study investigates the effect of TCE on neurogenesis using human induced pluripotent stem cell (iPSC) derived neural organoids to shed light on its neurotoxic effects in the developing human brain. We examined organoid, morphology, and gene expression when comparing control and TCE treated organoids. We found that in the TCE-exposed organoids, there was noticeable injury. Approximately 750 um in width, the scar contained an influx of astrocytes. The average diameter of the controlled organoids was 2.72 mm, whereas the average for TCE-exposed was only 2.27 mm. The significance of our results is that they demonstrate the impact of TCE on the developing brain.

Thi Le

Stanford

Mentor: Hope Lancero PhD

PI: Rosa Bacchetta MD

Poster #071

Poster #071

Title

Gene Expression of FOXP3 Isoforms in CD4+ T-Cells

Abstract

Forkhead box protein 3 (FOXP3) is the master transcription factor in Regulatory T cells (Tregs), a subset of CD4+ T cells. There are two predominant FOXP3 isoforms due to alternative splicing: a full-length isoform (FOXP3FL) and one lacking exon 2 (FOXP3∆2). FOXP3 gene mutations lead to a loss of functional Tregs, leading to autoimmune diseases like IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X-linked) syndrome. We hypothesize that FOXP3FL gene expression will be more present than the FOXP3∆2 gene expression upon cell activation in healthy human CD4+ T cells. We isolated RNA from activated CD4+ T cells and utilized reverse-transcription polymerase chain reaction to quantify FOXP3FL and FOXP3∆2 isoforms. The full length isoform was more abundant than the FOXP3∆2 isoform and FOXP3∆2 decreased over the activation course of 8 days. These findings demonstrate that substantial FOXP3FL is produced upon activation of CD4+ T cells at the mRNA level.

Jacky Law

Stanford

Mentor: Yuan Guan, PhD

PI: Gary Peltz MD/PhD

Poster #072

Poster #072

Title

Investigating Vorinostat’s Effect on Liver Fibrosis

Abstract

Liver fibrosis is a life-threatening disease that occurs in response to chronic liver injury characterized by the accumulation of extracellular matrix (ECM), which includes fibril-forming collagens. Chronic liver injury causes hepatic stellate cells to transdifferentiate into myofibroblasts, which secrete fibril-forming collagens and other ECM proteins causing liver fibrosis. As fibrosis develops, blood flow within the liver is restricted, hepatocytes die, and the liver cannot perform its essential functions. In this project, we generated fibrotic human Hepatic Organoids (HOs) (supplemented with the growth factor TGF-β1) from induced pluripotent stem (iPS) cells and investigated the effects of FDA-approved histone deacetylase inhibitor Vorinostat (SAHA) on fibrosis. Interestingly, we found that it reduced the amount of collagen (COL1A1) expressed, which suggests that Vorinostat could be an effective clinical tool for treating liver fibrosis.

Yunxi Zhang

Stanford

Mentor: Kensuke Kayamori MD/PhD

PI: Ravi Majeti MD/PhD

Poster #073

Poster #073

Title

HOPX effects on Acute Myeloid Leukemia Growth Rate and Chemosensitivity

Abstract

The gene HOPX is highly expressed in leukemia stem cells but lowly in leukemia progenitor cells1, and its high expression correlates with poor prognosis in AML patients2. Investigating the role of HOPX expression in the KG-1 AML cell line seeks a more effective AML therapeutic target. We hypothesized that HOPX knock-out (KO) in KG-1 cells would influence their growth rate and chemosensitivity. We utilized the homology-directed repair gene-editing method to delete HOPX in KG-1 cells. By tracking the growth of KG-1 Control and HOPX KO cells for 15 days, we found both groups exhibited exponential growth, but cell numbers were continually similar. The chemotherapeutic agent Cytarabine was added to another set of samples to test chemosensitivity. Viable cell numbers of the two groups demonstrated no significant difference after 13 days of counting. These data reflect that HOPX expression has no statistically significant impact on KG-1 cells’ growth rate or chemosensitivity.

Thu Lei

Stanford

Mentor: Hope Lancero PhD

PI: Rosa Bacchetta MD

Poster #074

Poster #074

Title

Protein expression of FOXP3FL and FOXP3Δ2 isoforms in activated CD4+ TCells

Abstract

FOXP3 is the master transcription factor of regulatory T cells containing two isoforms in humans. However, the dynamic protein expression of these isoforms in CD4+ T cells after activation is not well known. The current study is to analyze the protein expression of two isoforms, FOXP3FL and FOXP3Δ2, after activating CD4+ T cells via T Cell Receptor (TCR) by anti-CD3/CD28 beads. The activated cells were collected at timepoints Day 3, 6, 7 and 8. Our aim was to analyze the dynamic changes of protein expression of both isoforms after activation. Protein expression was analyzed by Western Blots and quantified using ImageJ software. The results show that protein expression of both isoforms followed similar trends and peaked on days 3 and 7 after activation. This finding indicates that while they do play a role in protein expression, other components may influence expression patterns over time.

Diane Joung

Stanford

Mentors: Jinyi Xiang MD/PhD, Christopher Sun and Laura Almeida BS

PI: Irving Weissman MD

Poster #075

Poster #075

Title

Exploring FGFR3 Expression is Hematopoietic Stem Cells

Abstract

The hematopoietic system generates over 100 billion blood cells daily, essential for oxygen transport, immunity, and tissue repair. Hematopoietic stem cells (HSCs) drive hematopoiesis by continuously renewing, proliferating, and differentiating into various blood cells within the bone marrow (BM). This study focuses on the endosteal niche, located along the bone's inner surface, and explores the hypothesis that long term (LT) HSCs express the FGFR3 receptor in this niche. We utilized mouse perfusion, cryosectioning, immunofluorescence staining, and microscopy to investigate the expression of FGFR3 in Hoxb5+ LT-HSCs in whole bone marrow using our Hoxb5-tdTomato mouse model. We observed FGFR3 staining across LT-HSCs and other BM cells, suggesting a broader role across multiple niches and signaling pathways. Exploring FGFR3 regulation will enhance our understanding of FGF signaling in blood development, injury response, and cancer, potentially leading to new targets that govern HSC proliferation, migration, and metabolism, potentially improving therapeutic strategies.

Yuying Huang

Stanford

Mentor: Suyash Raj BA

PI: Irving Weissman MD

Poster #076

Poster #076

Title

Modeling Maternal Stress in a Dish: Cortisol Effects in Cortical Organoids

Abstract

Maternal stress during pregnancy is connected with an increase in stress hormone that affects the fetal brain. One mechanism of how maternal stress may impact the developing fetal brain is through the glucocorticoid hormone cortisol, which has previously been shown to increase levels of neuroinflammation. We hypothesized that cortisol will increase astrocyte activity by promoting the immune system’s response to injured central nervous system, which indicates higher neuroinflammation. To address our hypothesis, we generated cortical organoids (COs) from induced pluripotent stem cells (iPSCs) to examine the effect of cortisol on neuroinflammation during neural development. We dosed COs with either hydrocortisone or regular organoid growth medium for 2 weeks to stimulate stress. Using immunofluorescence, we found an increase in astrocyte and oligodendrocyte activity in the organoids with hydrocortisone treatment relative to controls.

Audrey Handoko

UC Riverside

Mentor: Mayra Daniela-Silva, Grad Student

PI: Dr. Huinan Liu

Poster #077

Poster #077

Title

Indirect Culture of the Cytocompatibility of Bone Marrow Stem Cells

Abstract

In this research project, the aim is to study the degradation of zinc alloys in vitro and to investigate different alloys that can be utilized in medical devices or implants to help treat conditions like osteoporosis. Zinc is used primarily due to its role in bone metabolism which promotes bone strength and formation. Compared to other elements such as Magnesium and Iron, it has moderate corrosion rates and does not produce harmful hydrogen ions. However, though it is less harmful, it is very brittle and not as resilient. To improve these properties, it can be alloyed with other metals like magnesium, calcium, manganese, and lithium to improve the strength and biocompatibility. In addition, this process also involves exposing the bio alloys to bone marrow stem cells (BMSCs) to understand the capabilities of these alloys. Bone marrow stem cells can help restore tissue that is damaged with individuals who have bone fractures. Overall, studying degradation of bio alloys and the cytocompatilbility with bone marrow stem cells will be able to help individuals who are diagnosed with osteoporosis and will repair or restore the damaged bone while reducing the risk of cytotoxicity.

Lucia Otanez

UC Riverside

Mentor: Vala Masjedizadeh PhD Student

PI: Xiaoping Hu PhD

Poster #078

Poster #078

Title

Bacterial Gene Verification (mms6) in Mammalian Cells

Abstract

Magnetic Resonance Imaging (MRI) has revolutionized medical diagnostics with its non-invasive, high-resolution imaging capabilities. Recent advancements integrate MRI with magnetic nanoparticles, which can enhance imaging precision and therapeutic efficacy. The mms6 gene plays a critical role in this innovation by facilitating the synthesis of magnetite nanoparticles, which are essential for targeted MRI applications and magnetic hypothermia treatments. Magnetite nanoparticles, produced with the aid of mms6, are used to improve contrast in MRI scans, allowing for more detailed visualization of tumors and other pathological conditions.

The current work aims to analyze HT-29 cells– a human colorectal cancer cell line commonly used in research due to their well-characterized genetic profile. Once the HT-29 cells are engineered to express the mms6 gene, they gain the ability to produce magnetite nanoparticles more efficiently. The laboratory work focused on verifying if said HT-29 cells accurately expressed the mms6 gene with the help of q-PCR.

Santo Quintero

UC Riverside

Mentor: Natalie Luu (Graduate Student)

PI: Dr. Ke Du (Assistant Professor, Chemical and Environmental Engineering Department)

Poster #079

Poster #079

Title

Nanofluidic Polydimethylsiloxane Chip

Abstract

Pathogen detection can be an important step in controlling the spread of disease (Dong et al., 2022). Pathogen testing is a process that can be lengthy, require specialized equipment, and require trained personnel (Dong et al., 2022). Portable and inexpensive nanofluidic chips, however, can help with pathogen detection. In this project, a nanofluidic polydimethylsiloxane chip was made. A patterned silicon wafer, silanization solution, PDMS, and silicone elastomer curing agent were used in conjunction with vacuum chambers, an ultrasonic cleaner, a plasma gun, and other devices to create a nanofluidic chip. A test with red dye showed a chip was successfully fabricated. In the future, the chip can help stem cell transplants by detecting viruses. Additionally, the chip can be used for duplex and avocado sunblotch viroid detection with the help of a prepared solution, such as a sample that has had fluorescence attached and been amplified, and an image reader.

Crystal Alvarez

UC Riverside

Mentor: Kevin Dalsania, Jonah Damian and Varun Vemari

PI: Dr. Iman Noshadi

Poster #080

Poster #080

Title

Digital Light Processing (DLP) Bioprinting of PGSA

Abstract

Chronic or acute ligament injuries often have limited healing potential due to low
blood flow and limited cell density within the tissue. Tissue engineering is currently being
explored as a promising possible treatment to repair damaged ligaments but challenges
emerge to identify a scaffold material that can withstand the tensile forces in the ligament
while maintain high cell survivability. Additionally, each defect site will vary in shape or
size requiring an optimal solution to account for all three factors. Therefore, we propose a
Digital Light Processing (DLP) bio-ink, based on the bioinert Poly (Glycerol-co-Sebacate)
Acrylate (PGSA) polymer conjugated with the cellular adhesion agent Bio-ionic Liquid
Bicarbonate (BIL-BC) (Bio-PGSA). Our results demonstrate a highly optimized bio-ink
that allows for the creation of high-fidelity structures opening the avenue to regenerative
medicine applications for ligament damage.

Alayna Yared

UC Riverside

Mentor: Natalie Luu (Graduate Student)

PI: Dr. Ke Du (Assistant Professor, Chemical and Environmental Engineering Department)

Poster #081

Poster #081

Title

Holy Guacamole! ASBVd Detection With A Microfluidic Chip

Abstract

Avocado sunblotch viroid disease (ASBVd) affects avocado production and trade. The disease reduces fruit yield and quality, highlighting the need for infield disease detection methods with high specificity that can detect both asymptomatic and symptomatic samples from different parts of the tree. RT-LAMP, an amplification method that makes copies of a target sequence of RNA for imaging, shows promise as it uses less lab equipment compared to the gold standard PCR (another common amplification method). This is partially due to LAMP being isothermal as the entire process happens at one temperature while PCR requires thermal cycling. Using a microfluidic chip to quantify viral load, the goal of the experiment is to test the use of RT- LAMP on the digital chip. The results of our experiment shows that RT-LAMP can detect both asymptomatic and symptomatic samples and samples from different parts of the tree.

Vani Gupta

UC Riverside

Mentor: Esther E. Omaiye, PhD

PI: Prue Talbot, PhD

Poster #082

Poster #082

Title

Cellular Effects of Vaping Liquids on Airway Basal Stem Cells

Abstract

The cigarette industry has been evolving, with electronic cigarettes (e-cigarettes) becoming increasingly prevalent as an alternative to smoking. Concurrently, ultrasonic cigarettes (u-cigarettes) have emerged as popular tobacco products. The objective of our study was to evaluate the effect of electronic and ultrasonic vaping liquids on Airway Basal Stem Cells (ABSCs). The ABSCs were cultured in T-25 flasks in accordance with established protocols and were treated with six flavor variants. Two SURGE u-cigarette flavor variants (“Green Mint” and “Polar Mint”) and four JUUL e-cigarette flavor variants (“Crispy Menthol,” “Polar Menthol,” “Ruby Menthol,” and “Summer Menthol”) were assessed. Cellular effects of the vaping liquids on ABSCs were analyzed using fluorescent microscopy and MTT Assay. The findings revealed that both SURGE and JUUL vaping liquids hindered cellular metabolic activity, mitochondrial reductases, and led to a reduction in cell spreading. The results underline how both vaping liquids have detrimental effects on the airway epithelium.

Yunshu Zhang

UC Riverside

Mentor: Isabella Bagdasarian, Ph.D Candidate

PI: Dr. Joshua T Morgan, Ph.D.

Poster #083

Poster #083

Title

3D Culture and Genetic Tools for Contractile iPSC-Derived Muscle

Abstract

Skeletal muscle tissue, essential for movement and metabolism, presents significant challenges for in vitro studies due to its contractile nature. Thus, there is a need for robust methods to model skeletal muscle physiology. For this purpose, the project I am working on focuses on two aspects: developing OptoTGFBRs-expressing muscle cells to regulate cell fibrosis in vitro, and optimizing dopamine-treated porous sugar posts for induced pluripotent stem cells (iPSCs) culture. Transgenic OptoTGFBR was cloned into a lentiviral backbone to be transduced into iPSCs. Further, optimization of porous polydimethylsiloxane (PDMS) molds treated with polydopamine was conducted using sugar as a porogen. Successful molds were appropriately constructed and sterilized. In future studies, OptoTGFBRs allows for light-controlled modulation of TGF-beta signaling and the PDMS supports proper contraction. These tools enable more robust iPSC-based skeletal muscle models to be utilized for in vitro differentiation and monitoring.

Kavya Kadakia

UC Riverside

Mentors: Mayra D'Silva (Grad Student) and Yiqing Chen (Post Doctorate)

PI: Dr. Huinan Liu (Director of STRIDE program)

Poster #084

Poster #084

Title

Direct Culture and Cytocompatibility of Zinc Alloys in BMSCs using ICP

Abstract

As our population ages, so does the relevance of osteoporosis in our society. While implants have consistently proven their efficiency in “mending” broken bones, finding the right material for these implants that is biocompatible and cost efficient is a challenge. The study analyzes 3 compositions of zinc alloys in swine derived bone marrow mesenchymal stromal cells. Direct culture studies involve seeding cells on alloy samples in 12-well plates, then incubating the plates for 24 hours, and results quantify ion concentrations with ICP-OES. This is followed by fluorescence microscopy to quantify cytotoxicity, by fixing and staining adherent cells, and quantifying cell density, spreading and viability using fluorescence microscopy and ImageJ software. This study not only paves the way to promise material efficiency, but also to minimize future complications, offering a sustainable solution that could significantly enhance patient outcomes and reduce long term healthcare expenses.

Julian Methippara

UC Riverside

Mentor: Vinit Adani; Graduate Student

PI: Dr. Martin I. Garcia-Castro; PhD

Poster #085

Poster #085

Title

The Expression of GLI3 in Chick Cranial Neural Crest Cell Development

Abstract

Neural Crest Cells (NCCs) are important stem-like progenitors originating at the neural plate borders, induced by different signaling molecules like BMPs, FGFs, and WNT. These molecules activate transcription factors (e.g., Pax7, Msx1, and Zic) and initiate NCC development. NCCs go through a gene regulatory network and eventually delaminates from the neural tube and differentiate into different cell types like peripheral neurons, glia, bone, and cartilage. This study investigates GLI3, a gene in Sonic Hedgehog signaling pathway, which is responsible for organ and tissue patterning, including head and limbs. GLI3, a transcription factor, regulates gene expression by binding to specific regions of DNA. Preliminary data indicates GLI3 involvement in Cranial NCC gene regulatory network, however its function remains unknown. Results from In-situ hybridization show GLI3 expression in Cranial region of embryo, where Cranial NCC are formed and differentiate. Further studies are needed to assess GLI3’s role in Cranial NCC development.

Isabella Mocino

UC Riverside

Mentor: Iris Andrade

PI: Sonali Chaturvedi, Ph.D.

Poster #086

Poster #086

Title

The autorepression circuit in CMV dictates the fate of the virus

Abstract

Cytomegalovirus (CMV) is the leading cause of birth defects and transplant failure. More than 50% of transplant patients are infected with CMV and more than 70% of the global population is infected with it. CMV encodes an autorepression circuit (IE2), that regulates its own expression by binding to its promoter. Disruption of this IE2 autorepression circuit induces “open-loop lethality”, and reduces viral fitness, hence the autorepression circuit dictates the fate of the virus (Chaturvedi et al. Cell 2022). By PCR amplification of IE2 gene, its promoter and a reporter gene followed by assembly by gateway cloning will provide us with minimal feedback circuit, this circuit will be used to create a stable cell line using lentivirus in ARPE-19 cells. The goal of this project is to study how CMV evolves in ever changing environments and the mechanism of autorepression circuits dictates viral fitness.

Micah Vo

UC Riverside

Mentor: Aihik Banerjee. Grad Student and Kevin Dalsania. Grad Student

PI: Dr. Iman Noshadi

Poster #088

Poster #088

Title

3D Optical Bio Printing for Tissue Regeneration

Abstract

Biomaterials and their functionality require optimization techniques that enhance viability with cells and tissue for cardiac regenerative applications. While there is an abundance of biomaterials, there remains an absence of advanced techniques, creating restrictions and issues on addressing cardiovascular problems. In response to this, tissue engineering through 3D optical printing has developed in the field to produce structures compatible with the heart. In Dr. Noshadi’s lab, we developed a photocrosslinked scaffold through use of a digital light processing bioprinter utilizing fish biogel. The optimal structure was determined through optimizing the bioprinter’s light, creating the best structure that was not under polymerized nor over polymerized. The assessment of the printed structures showed compatibility with cells, demonstrating the potential of optimizing biomaterials 3D optical printing techniques in cardiovascular regenerative applications.

Charynna Pe Benito

UC Davis

Mentor: Casiana Gonzalez, PhD

PI: Kyle Fink, PhD

Poster #093

Poster #093

Title

Characterizing and Validating CRISPR/dCas9 iPSCs to model CASK disorders

Abstract

Mutations in the calcium/calmodulin dependent protein kinase gene (CASK) are responsible for X-linked CASK-related disorders affecting neurodevelopment. This study aims to compensate for the current lack of model systems for CASK-related disorders by creating a disease-relevant model. Our approach uses CRISPR/dCas9-edited induced pluripotent stem cells (iPSCs) with a four base-pair deletion to knock out CASK. Validation of the iPSCs was conducted to ensure appropriate CASK expression in the cell lines and characterization to assess pluripotency. Pluripotency was evaluated through immunocytochemistry (ICC) and sequences were confirmed via gDNA and RNA sequencing. The iPSCs demonstrated positive pluripotency markers, however, the CASK marker could not be visualized. This indicates the need for further validation. Future objectives will involve differentiating the iPSCs into neurons for therapeutic testing. A promising approach to restore CASK gene function is transcriptional upregulation and promoter demethylation of the XCI-silenced WT CASK allele for a rescue.

Chigozie Monye

UC Davis

Mentor: Angelica Bachman, MA

PI: Jan Nolta, PhD

Poster #094

Poster #094

Title

Manufacture of lentivirus to model Fibrous Dysplasia using Mesenchymal Stro

Abstract

Fibrous dysplasia is a rare genetic disease resulting in scar-like tissues instead of healthy bone. It’s a sporadic non-hereditary disorder caused by a gain of function mutation in the GNAS gene that encodes for Gsɑ protein. This mutation can lead to fibrous tissues growing in place of healthy bone, making bones weak and brittle over time. We aimed to package lentivirus coding for the mutated and wildtype gene to model both healthy and fibrous dysplasia cell types.. We packaged the wildtype and mutant gene into a lentivirus using HEK-293T cells to model this disease in a cell. We then used human bone marrow-derived mesenchymal stromal cells (MSCs) to validate the virus's ability to transduce our cells of interest. Our findings revealed that both viruses successfully transduced MSCs, integrating the wildtype and mutant genes. This means the virus was successfully manufactured and can be utilized for future studies.

Paul Calmac

UC Davis

Mentor: Ping Zhou, PhD

PI: Ping Zhou, PhD

Poster #095

Poster #095

Title

How Cri du Chat affects CTNND2 and SEMA5A in iPSCs vs. NPCs

Abstract

Cri du Chat Syndrome (CdC) is a rare genetic disorder that leads to facial deformities and intellectual disability. It is caused by a deletion of one of the 5p chromosomes, including the CTNND2 and SEMA5A genes. The goal of this study is to assess whether the expression CTNND2 and SEMA5A are impaired in the patient's neuronal cells. We obtained induced pluripotent stem cells (iPSCs) from both healthy and CdC-affected individuals and then differentiated these iPSCs into neural progenitor cells (NPCs). In healthy patient cells, qPCR analysis showed that CTNND2 was expressed significantly more in NPCs than in iPSCs. However, the NPCs derived from a CdC patient showed less expression of the CTNND2 gene than their iPSC counterpart. Immunostaining assay further confirmed delta-catenin was also produced in the healthy NPC cells more than that of the disease-affected NPCs. However, the diseased NPCs did not show any reduction in SEMA5A expression.

Yuvraj Gill

UC Davis

Mentor: Atul Dave, PhD

PI: Maneesh Dave, MD

Poster #096

Poster #096

Title

Human MSCs modulate SAMP macrophages to anti-inflammatory phenotype.

Abstract

Crohn's Disease (CD) is characterized by overactivation of the human immune system. Currently available therapies show promising results, however, disease relapse is still very common in CD patients. Mesenchymal Stem Cells (MSCs) therapy have shown great potential in immune related diseases such as CD. Recently, Dave et al. (npj Regen Med,2024) have shown that human MSCs are immunosuppressive and have long-term efficacy in SAMP mice, a spontaneous murine model of small intestine inflammation resembling human CD. Single cell RNA sequencing of SAMP mesenteric macrophages (MФ) at day 28 of the MSCs treatment have shown that MSCs had sustained effect via downregulation of CDk8 and upregulation of Gas7 genes in mesenteric MФ. Here, we performed an in-vitro co-culture assay with human MSCs and LPS induced SAMP peritoneal MФ. We observed that MSCs downregulate expression of Cdk8 gene, along with pro-inflammatory cytokines TNF-α, IL-1β and upregulate anti-inflammatory marker Arginase-I in MФ.

Angelina Gan

UC Davis

Mentors: Nik Lind, PhD and Anber Sitara Ansari, MS

PI: Julian Halmai, PhD

Poster #097

Poster #097

Title

Comparative Analysis of CMV Against SCP in SMC1A Expression Regulation

Abstract

Loss-of-function mutations in SMC1A result in SMC1A-related Developmental and Epileptic Encephalopathy (SMC1A-DEE) which is a severe neurodevelopmental disorder characterized by intractable seizures, developmental impairment, intellectual disability, and other neurological abnormalities. Through traditional gene therapy, we can reintroduce SMC1A and improve the outlook for patients with SMC1A-DEE. While this method presents a great opportunity to be utilized, large promoters exceed the packaging capacity of Adeno-Associated Virus, the current gold standard for gene supplementation strategies, prompting the need for smaller promoters for SMC1A delivery. Our solution is to replace the CMV promoter with a 87-bp super core promoter (SCP). We tested SCP-SMC1A in HEK293 and found that there was a significant decrease in SMC1A gene expression when compared to the CMV promoter. These findings illustrate the potential of using SCP for future gene supplementation of additional targets, potentially presenting a solution for the treatment of other loss-of-function mutation diseases.

Manahil Rehman

UC Davis

Mentor: Amin Cressman, MS

PI: Fernando Fierro, PhD

Poster #098

Poster #098

Title

Genetic Engineering of SSCs to Promote Their Expansion In Vitro

Abstract

Skeletal stem cells (SSCs) have great potential as a regenerative therapeutic for skeletal disorders such as osteoporosis.One drawback is the current inability to expand these cells in vitro without undergoing differentiation or senescence. Telomeres are repeating DNA sequences flanking eukaryotic chromosomes that provide genomic stability.Telomerase (TERT), a reverse transcriptase activated in stem and progenitor cells, rebuilds these ends as they are shortened after repeated cell division.We hypothesize that transiently overexpressing TERT in SSCs will counteract telomere shortening and promote proliferation, allowing for in vitro cell expansion. To achieve this, human TERT was ligated into a pCCLc cloning vector to be taken up by competent E.coli cells.The recombinant plasmid will be used to transfect HEK293T lentiviral packaging cells, and the harvested lentivirus will be used to transduce SSCs, overexpressing TERT.The in vitro expansion of SSCs enhances the potential for regenerative therapeutics targeted towards skeletal defects.

Kelly Loi

UC Davis

Mentors: Ryan N. Nielsen and Michael K. Sheng

PI: William J. Murphy, Ph.D.

Poster #099

Poster #099

Title

Comparing CAR T cell Expansion and Phenotype Using Different Culture Medium

Abstract

CAR T-cell therapy is a novel treatment where T cells are isolated from a patient and genetically engineered to detect and eliminate cancer cells. Clinically, they have led to astounding success in treating many B cell cancers, serving as a lifesaving option for patients who fail to respond to conventional therapies such as radiation and chemotherapy. However, researchers are still attempting to optimize culture conditions to improve CAR T cell expansion and efficacy. In this study, we generated and cultured CAR T cells using two commercially available media: TexMACS and X-Vivo 15, to measure cell expansion and phenotype via flow cytometry. Results indicate that CAR T cells cultured in X-Vivo 15 exhibit significantly increased expansion and a higher prevalence of CD8+ T cells compared to those cultured in TexMACS, suggesting X-Vivo 15 may be a more suitable medium for CAR T cell production.

Sanford Burnham Prebys Medical Discovery Institute

Mentors: Sofia Lombardi, Bachelor of Science, Research Assistant and

Meenakshi Sudhakaran, PhD, Postdoctoral Associate

PI: Kelly Kersten, PhD

Poster #122

Poster #122

Title

Spatial Organization of Exhausted CD8+ T-Cells in Melanoma Skin Cancer

Abstract

Melanoma, an aggressive type of skin cancer, is characterized by a significant infiltration of immune cells. Understanding the cancer-immune interplay in the tumor microenvironment (TME) is imperative for developing effective immunotherapies. Anti-tumor CD8+ T cells in the TME often become exhausted and dysfunctional due to chronic antigen exposure, resulting in high PD1 expression and reduced cytotoxicity. We hypothesized that exhausted CD8+ T cells reside in specific niches in the TME that support interactions with other immune cells such as macrophages, contributing to immune suppression. We used immunofluorescence (on cryosections) to study the localization of PD1+ CD8+ T cells in relation to CD31+ blood vessels, and CD11b+ macrophages in mouse spleen and B78ChOVA melanomas. We found that CD8+ T-cell exhaustion is significantly more pronounced in the TME compared to the spleen. Future studies to understand the detailed spatial distribution of immune cell neighborhoods in the TME should be pursued.

Jason Zhang

Sanford Burnham Prebys Medical Discovery Institute

Mentors: Erik Hultenius, PhD student and Mehran Ghafari, PhD

PI: Xiao Tian, PhD

Poster #123

Poster #123

Title

Astrocyte and microglia isolation to investigate neuroinflammation in aging

Abstract

The Information Theory of Aging states that aging is driven by a gradual loss of epigenetic information. Consistent with the Theory, epigenetic changes are at least partially responsible for increased neuroinflammation, a primary driver of the brain aging phenotype. For example, microglia are known to both accumulate and deteriorate with age while promoting inflammation. Aged astrocytes, like microglia, also experience moderate heterogeneous inflammation and dysfunction, leading to synapse elimination. The aging of the central nervous system thus is a main contributor to the decreased cognitive function seen with age. With this in mind, we attempt to establish an in-vitro system to identify markers of young and old glia through the culture of primary cells and to develop a sophisticated image-analysis method to differentiate between young and old cells. Such a system is crucial in validating potential epigenetic interventions(e.g. rejuvenation) that may target neuroinflammation and the underlying aging mechanism.

Naomi Shim

Sanford Burnham Prebys Medical Discovery Institute

Mentors: Luca Caputo, PhD, and Monica Nicolau

PI: Pier Lorenzo Puri, MD

Poster #124

Poster #124

Title

Uncovering the role of the lncRNA EPR within the human intestine

Abstract

LncRNAs, defined as RNA molecules >200nt without protein coding capabilities, possess a critical role in gene regulation, both in physiological (e.g., development) and pathological processes (e.g., cancer). Previous studies in mouse models have discovered EPR as a key regulator of gene expression and as a tumor suppressor in both mammary glands and intestines. Bioinformatic analysis of RNA-seq in a colon-specific EPR-KO mouse model uncovered a novel role in regulating the MAPK signaling pathway. In this project, we investigate the role of EPR within the human intestine. In order to understand EPR’s role and regulation in human tissue, we have cloned its putative promoter to develop luciferase vectors and generated intestinal organoids from healthy donor hiPSCs. Initial expression analysis revealed EPR expression in late-stage organoids, suggesting that EPR is dispensable during human intestine development, but may potentially play a role in regulating gene expression in fully differentiated tissue.

Nellie Perez

Sanford Burnham Prebys Medical Discovery Institute

Mentor: Sophie Hao, B.S.

PI: Duc Dong, Ph.D.

Poster #125

Poster #125

Title

Investigating the Extrahepatic Duct as a Source of Liver Regeneration

Abstract

The liver is a highly regenerative organ, but regenerative properties have not been observed in patients with genetic diseases like Alagille Syndrome (ALGS), characterized by the absence of intrahepatic ducts (IHDs) due to a defective Notch pathway. As a result, bile is unable to be transported to the gallbladder, causing liver failure. Previously, we showed extrahepatic ducts (EHD) housed liver stem cells that led to IHD regeneration in a zebrafish ALGS model. Another liver disease, Biliary Atresia, primarily affects EHD and causes patients to develop intrahepatic pathologies later in life. We aim to investigate EHD's role in liver regeneration and apply our findings to human liver disease patients. We are utilizing CRISPR-Cas9 to generate a novel lineage tracing line labeling in the EHD and establishing liver damage models in zebrafish. The findings will inform the role of EHD in liver regeneration and potentially therapeutic strategies targeting this liver stem cell source.

Sara Hansali

Sanford Burnham Prebys Medical Discovery Institute

Mentor: Alicia Llorente Lope, PhD

PI: Brooke M. Emerling, PhD

Poster #126

Poster #126

Title

The Role of PI5P4Ky on Targeted Therapy Resistance in HER2+ Breast Cancer

Abstract

HER2+ breast cancer accounts for ~30% of total breast cancer diagnoses and is characterized by amplification of the ERBB2 gene, leading to over expression of HER2 receptors on the plasma membrane. Although the HER2+ subtype is considered an aggressive subtype, patients typically respond to targeted therapies such as Trastuzumab and Lapatinib in early stages. However, the development of treatment resistance over time can lead to recurrence and metastasis, which highlights the need for new strategies to overcome anti-HER2+ drug resistance. We have identified a link between elevated expression of the lipid kinase PI5P4Kg and poor outcomes in HER2+ breast cancer patients. In this study, we have generated and validated several in vitro models of acquired resistance to Trastuzumab and Lapatinib to investigate the role of PI5P4Kg in the development of resistance to anti-HER2 therapies.

Saanvi Dogra

Sanford Burnham Prebys Medical Discovery Institute

Mentor: Diana Presas

PI: Sanjeev Ranade

Poster #127

Poster #127

Title

Streamlined Platform for Predicting Pathogenicity of Genetic Variants

Abstract

Congenital heart disease (CHD) is the most common form of birth defects. CHD is often caused by combinations of variants affecting multiple proteins; however, defining the effects of genetic variants on protein function remains a major challenge. Here, we developed an algorithm to web scrape pathogenicity scores from an artificial intelligence tool, AlphaMissense, and integrate these scores with previously developed tools using Ensembl Genome Browser. We correlated AlphaMissense predictions against Ensembl scores for GATA4, an essential transcription factor in cardiac development. We find that pathogenic variants predicted by AlphaMissense and Ensembl were concentrated near the DNA binding zinc-finger domains. In total, however, our algorithm found nominal cross-correlation (R2 = 0.49) in GATA4 variants, indicating a lack of reliability when using only a single predictor as a basis for scientific experimentation. For future efforts, our streamlined algorithm can be broadly applied to determine pathogenicity of protein coding variants in CHD cases.

Billie Joelle Bote

Sanford Burnham Prebys Medical Discovery Institute

Mentor: Dr. Deron Herr

PI: Dr. Jerold Chun

Poster #128

Poster #128

Title

Somatic Genomic Recombination in Alzheimer’s Disease

Abstract

Alzheimer’s Disease is the most common cause of Dementia. It is a progressive disease that involves the buildup of Amyloid-𝛽 plaques in the brain. Previous work done by the Chun Lab found that a possible cause of the Amyloid build-up is Somatic Genomic Recombination, in which an insertion mutation is caused by endogenous viral-like elements. This reuslts in a new DNA Sequence known as a “gencDNA.” Our hypothesis is that Polymerase Chain Reaction (PCR) can be used to estimate the frequency of gencDNA formation for specific genes in the mouse brain. We have performed PCR for 3 different genes using mouse brain cell lines with known gencDNAs as controls. After validating the methodology, we performed PCR on Mouse Prefrontal Cortex DNA to see how often each specific gencDNA occurs and have determined that the evaluated genes form gencDNAs approximately once per 10,000 cells.

Shawn Delaware

Sanford Burnham Prebys Medical Discovery Institute

Mentors: Gabriele Guarnaccia, BSc, MSc and Collin Kaufman, PhD

PI: Alessandra Sacco, PhD

Poster #129

Poster #129

Title

The Effects of Cachexia During Muscle Differentiation

Abstract

Skeletal muscle is a voluntarily-controlled tissue necessary for breathing and locomotion. Skeletal muscle self-regenerates after injury through a differentiation process called myogenesis, which is impaired by diseases like cancer cachexia, a progressive muscle mass loss associated with cancer. How cancer cachexia negatively affects muscle is poorly understood, hence the design of models to investigate this mechanism is a major unmet need. To achieve this aim I tested, in vitro increasing seeding densities (5,000-30,000) for optimal C2C12 myoblast differentiation. Immunostaining revealed an optimal plating density of 62,500 cells/cm2. To mimic cachexia in vitro, I applied conditioned media (CM) from KPC-Pdx1 cancer cells to C2C12s. CM resulted in a reduced myotube diameter, revealing decreased differentiation and cell fusion. This in vitro model will help define the role of cachectic factors in myogenesis. My project will provide a better understanding of the molecules causing cancer cachexia, which will lay the grounds for cancer cachexia therapies, thereby improving cancer patients’ life expectancy.

Elena Salcedo

Sanford Burnham Prebys Medical Discovery Institute

Mentor: Sara Ancel, Ph. D

PI: Yu Xin, Ph. D

Poster #130

Poster #130

Title

Analyzing Glycosylation patterns in GNE myopathy using GLYPH and AI models

Abstract

GNE myopathy is a rare autosomal recessive muscle disorder caused by mutations in the GNE gene. GNE encodes an enzyme crucial for sialic acid production in surface sialyation, a glycosylation modification. Current imaging technologies struggle to detect glycosylation effectively. Thus, our lab developed GLYPH (Glycosylation Landscape analysis by Probe Hybridization) which can visualize proteins and Glycosylation moieties by multiplexed image. Using GLYPH, we examined 40 lectins in control and GNE biopsies, revealing widespread glycosylation changes indicating drastic impacts from the GNE mutation and normal degeneration. AI-assisted analysis of muscle biopsies showed a decrease in healthy fiber percentage with disease severity, alongside unexpected myofiber size increases with progression. Increased PTA galactose foci in the ECM and WGA signals correlating with fibrotic accumulation suggest potentially elevated compensatory glycosylation, counter intuitive to GNE function understanding. This warranting further investigation into lectin binding to extract the precise glycosylation changes.

Ava Teng

Colin Hsiang-Hsien Hwang

Daniel Svediani

​Ethan Sung

​Irene Thomas

Keith Mahar

Madison Cheungsomboune

​Sarah Martellino

Selin Bilgi

​Urja Parekh

Samaria Bailey

Brianna Boye

Emma Do

Kenneth Garnett

Brian Higgins

Anaiya Mayfield

Nicolas Mendoza

Estephanie Salinas

EwongoAbasi Umoh

Charlie Perez

Bianca Chirrick

Bernice Nkemnji

Isabella Martinez

Allison Paredes

Jennifer Ramirez Flores

Serena Hannifin

Isabella Krauss

Kaitlyn Morrison

Anna Villatoro

Seth Pickens

Obianuju Oyeoka

Kristy Xiao

​Nicole Padilla Alvarado

​Nghi Nham

Belem Osorio

Mercy Niyi-Awolesi

​Vy Phan

​Katherine Bien

Isaac Jung

Jonathan Li

Christine Lin

Jessica Lu

Daniel Nam

Natalie Sandoval

Jordan Struve

Julier Thein

​Angelina Wu

Aaron Yang

Alenabell Yang

​Vivian Yuan

Kira Zeng

Dyllan Fernandez

Joannah Jugalbot

Kimberly Vincent

Victor Martinez

Brook Zerihun

Jamie Ramos

​Jonathan Methippara

Brian Chen

Alberto Alonzo Garcia Brito

Saleh Saleh

Thi Le

Jacky Law

Yunxi Zhang

Thu Lei

Diane Joung

Yuying Huang

Audrey Handoko

​Lucia Otanez

Santo Quintero

Crystal Alvarez

Alayna Yared

​Vani Gupta

Yunshu Zhang

​​Kavya Kadakia

​Julian Methippara

​Isabella Mocino

Micah Vo

Charynna Pe Benito

Chigozie Monye

Ethan Sung

Irene Thomas

Sarah Martellino

Urja Parekh

Nicole Padilla Alvarado

Nghi Nham

Vy Phan

Katherine Bien

Angelina Wu

Vivian Yuan

Jonathan Methippara

Lucia Otanez

Vani Gupta

Kavya Kadakia

Julian Methippara

Isabella Mocino

Michaiah Pettway

Amy Yu

Ingrid Regalado

Giselle Ornelas

Jennifer Liu

Daniela Galvan

Abner Martinez

Kevin Tran

Sophie Ayma

Sara Hansali

Saanvi Dogra

Billie Joelle Bote

Shawn Delaware