First Week; Shree Reddy

Hello! My name is Shree Reddy, and I am a recent graduate of Santiago High School in Corona, California, as well as an incoming freshman at Stanford University. This summer, I will be interning with the CIRM SPARK program, under my mentor, Dr. Saba Shahin.

Our lab applies gene manipulation and stem cell-based therapies for retinal degeneration. We inject stem cells in pre-clinical animal models such as retinitis pigmentosa and glaucoma rodent models to investigate the effects of treatment in rescuing different retinal cell populations, specifically the photoreceptors and RGCs. The fundamental aims of our lab research basically focus on rescuing the photoreceptor/RGC populations to restore as well as improve vision.

We perform functional tests like electroretinography [ERG], optokinetic responses [OKR], and optical coherence tomography [OCT] to check the response of photoreceptors and the RGCs. We also apply different techniques including histology, immunohistochemistry and immunofluorescence, qPCR, western blot as well as various multi-omics approaches viz., single cell RNA sequencing, proteomics and spatial transcriptomics. Using these techniques and approaches, we decipher the molecular underpinnings and underlying mechanisms in diseased retina and the changes in transcriptomic and proteomic landscape following treatment.

In this program, I hope to learn the fundamentals and basics of retinal substructures and functionality as well as about stem cell therapies in general. I look forward to working with my mentor and learning as much as I can through this amazing opportunity!

First Week; Anton Wagner

Hello, my name is Anton Wagner, and this is my first blog post in my role as stem cell research intern at the Cedars-Sinai Institute of Regenerative Medicine within the CIRM SPARK program, which is being executed in a virtual format due to Covid-19 restrictions. For almost 8 weeks, I’ll be a proud member of the Sheyn Lab, absorbing knowledge and working on a cutting-edge research project under the guidance of accomplished PI Dr. Dmitriy Sheyn and my brilliant mentor, Dr. Wensen Jiang.

As a rising senior at Crescenta Valley High School, equipped with completion in AP Biology, AP Calculus, AP Physics, Honors Chemistry and Programming, and with big dreams of a career in biomedical science, I thought I may have what it takes to intern in a sophisticated research lab. However, I did not feel confident when I came face-to-face with the outstanding scientists that have taken me, merely a High School student, on as an intern. My emotions have been a bit of a rollercoaster during this first week. The Cedars-Sinai onboarding process can be intimidating (as any start in a large organization) but everybody treated me with respect and made me feel like a highly valued new employee (although I am not an employee but an intern). Then, on the first day of science lectures and big introductions, I felt embarrassed when I was unable to fix the failure of my mic or the repeated loss of connection to MS Teams, the app Cedars-Sinai uses for virtual collaboration. But this incident made me experience right away that all these brilliant minds belong to down-to-earth people, very understanding, approachable, collaborative-minded, and helpful people that not only inspire me with their abilities and dedication to their research mission but they truly accept me because they are committed to developing new generations of scientists.

During our first meeting, Dr. Sheyn and Dr. Jiang gave me a great overview of their work and my own assignment that they thoughtfully designed to focus on genomics-based analytics since I do not have access to perform wet lab tasks. For the rest of the week, my lab leaders have always made time to exchange with me and ease me into the significance, innovation, objectives, methods and processes of the research I will be involved in.  The work of the Sheyn Lab revolves around addressing a major health problem that affects millions of people, namely, back pain and mobility problems due to the degeneration of intervertebral discs. Intervertebral discs, consisting of a fibrocartilage tissue, function as a cushion and ligament between the vertebrae in the spine. Although intervertebral disc disease is common, the mechanisms of development are not well-understood, and there is a lack of clinical therapy. Stem cells with their fascinating properties of assuming differentiated cell functions have great potential to solve the therapy gap. However, in order to direct the stem cells to promote disc rejuvenation, the gene-regulated cellular responses of the disc cells, such as gene expressions and transcription factors, need to be better understood. To this end, the research team has performed single cell RNA sequencing of patient sample groupings. This is where my assignment comes into play which includes gene ontology data evaluation, deduction of the regulatory network, and scientific plotting. While writing this, I realize how many new insights I have already gained within the last few days by immersing myself in research review papers and interacting with Dr. Sheyn and Dr. Jiang. With excitement, I look forward to next week to deepen my conceptual knowledge and develop my analytical skills.

First Week; Kasey Afshani

Hi! My name is Kasey Afshani and I’m a part of the RMI CIRM SPARK program at Cedars Sinai this summer! I’m a rising senior at The Buckley School and have been given the amazing opportunity to work in the Svendsen Laboratory under my mentor, Dr. Deepti Lall. We are studying amyotrophic lateral sclerosis, more commonly known as ALS or Lou Gherig’s Disease. ALS is a fatal neurodegenerative disorder that affects the upper and lower motor neurons of the body, leading to paralysis and death within 3-5 years of diagnosis. Patients diagnosed with ALS present motor dysfunction such as muscle weakness and inability to move arms and legs due to the loss of neurons that control these movements. Unfortunately, there are no treatments available to cure this disease.

Last week, one of the only things I knew about ALS was that an ice bucket challenge to promote awareness surrounding ALS went viral a few years ago. This week, Dr. Lall has exposed me to several scientific research articles that have blown my mind wide open. At the start of the program, I felt intimidated by the vast complexity of the workings and studies of ALS, but now, under the guidance of Dr. Lall, I feel that I’ve begun to grasp the inner workings of the brain.

My research will center around the disease mechanisms for ALS and possible therapeutic approaches for the disease. So far, I’ve learned about different ways that a mutant protein can lead to ALS, one of those proteins being SOD1. Due to structural instability, the enzyme can misfold and accumulate in the motor neurons in the central nervous system. This disrupts mitochondrial function and calcium homeostasis, which leads to energy depletion and can lead to synaptic dysfunction and a loss of neurons.

I knew that I was always fascinated by the brain, but my interest in biomedical research began three years ago when I befriended someone who was diagnosed with diabetes mellitus when he was ten. I took to the internet to try and understand the ways in which diabetes not only inhibited the production of insulin, but why alpha cells never produced glucagon either. Sadly, I wasn’t well-versed enough in researching to understand enough.

However, I did not have the same guidance that I have at CIRM SPARK. Dr. Lall has given me the resources and the encouragement to read and understand scientific research papers in a way that I’ve never been able to before. I am truly thankful for her taking time out of her busy schedule to mentor me. I’m thrilled to learn all about ALS for the next six weeks!

First Week; Angela Lee

Hello! My name is Angela Lee and I am a CIRM SPARK intern at Cedars-Sinai. I am currently a rising senior at Milpitas High School in Milpitas, California. This summer of 2021, I am working at the Ljubimov Lab of the Regenerative Medicine Institute (RMI) under the guidance of my mentor Dr. Ruchi Shah.

On the first day, I was introduced to stem cell research by Dr. Wafa Tawackoli and learned about sterile techniques and safety procedures in labs. It was interesting hearing about the importance of stem cell research and its potential for treating diseases such as diabetes and heart disease. Stem cells provide the potential ability to grow new cells in a lab to replace damaged tissues or organs and accelerate healing through regeneration.

On my second and third day, my mentor Dr. Ruchi Shah provided me with an overview of the Ljubimov Lab. She briefed me on the lab’s extensive research on limbal epithelial stem cell research concerning epithelial wound healing in diabetic corneas. I learned about diabetic eye diseases, including diabetic retinopathy, and the role of limbal epithelial stem cells (LESC) in regeneration for new corneal epithelium. Dr. Shah and I looked at the comparison of epithelial wound healing in diabetic and non-diabetic corneas. Dr. Shah shared about the benefits of Wnt5a in corneal wound healing. One of the goals in this research is to find a way to increase Wnt5a protein in the diabetic LESC through gene therapy instead of having to add Wnt5a protein from an external source. The overall goal of the research being done in this lab is to find new ways to accelerate and improve diabetic wound healing.

By the end of my first week, my knowledge of scientific concepts and methods has increased substantially. I have been taught challenging terms such as diabetic retinopathy and LESCs, and have also learned multiple lab techniques. While I have previously only learned through textbook readings and minor lab assignments at school, Cedars-Sinai has provided me the tools and resources to further explore this innovative field of research and expand on my previous knowledge. I was given further insight on the use and impact of different concepts I have briefly learned in high school courses, including epigenetics and the use of antigen and antibody interaction to detect marker expression in  different cells. In the next seven weeks to come, I hope to be able to contribute data to this research by performing my own procedures in identifying changes in protein expression that will give us insight into the contrasts of wound healing between normal and diabetic corneas.

First Week; Maxwell Yao

Hi everyone! My name is Maxwell Yao. I just graduated from Canyon Crest Academy in San Diego, and I will be a freshman at the University of California, Los Angeles this coming fall. I’m very grateful to have been given the opportunity to experience professional research as a CIRM SPARK intern here at Cedars-Sinai, and my primary goal is to learn as much as I can about regenerative medicine so that I, too, can help people through scientific research.

This summer, I’m lucky enough to be able to work with Dr. David Rincón Fernandez Pacheco, a postdoctoral researcher of the Breunig Lab. He did his PhD in biomedicine in 2016 at the University of Córdoba in Spain and has been with Cedars-Sinai since 2017. His current research focuses on analysis of pediatric brain tumors through a murine model developed by the lab.

Even though it has only been a few days, I’ve already learned so much from Dr. Rincón Fernandez Pacheco. A majority of pediatric brain cancers contain mutations of the K27 or G34 types on the H3F3A gene, both of which are mutually exclusive and lead to the development of extremely different brain tumors. I also learned about organoids (structures grown from stem cells, consisting of organ-specific cell types that grow in a 3D, spherical shape). It’s amazing how these structures can mimic characteristics of actual tumors which allows for us to expand our understanding of cancer and develop more effective, personalized treatments for patients.

Additionally, Dr. Rincón Fernandez Pacheco taught me about the process for making the brain organoids we will be studying. Through electroporation after a DNA plasmid injection into the ventricles of a newborn mouse, a dual recombinase process edits the Rosa26 mtmG gene of the mouse and substitutes a certain section of that gene with the specific oncogenes we will be using (i.e TP53, PDGFRA and either the K27M or G34R/V oncogene variants). After allowing the tumor to grow in the mice, they are harvested which enables us to culture them as brain tumor organoids in the lab for analysis. I also got to watch my mentor give the organoids fresh, new media and cut the organoids with a tissue slicer, which was very fascinating.

Throughout this first week, my experience has been exceedingly positive and Dr. Rincón Fernandez Pacheco has been really helpful, teaching me the ins and outs of research, sending me background information, and answering whatever questions I may have. With such an exciting first week, I really look forward to learning more and more about regenerative medicine while simultaneously getting a glimpse into the world of professional medical research.

First Week; Eamon Bashiri

Hi, my name is Eamon Bashiri. I am a recent graduate from Oak Park High School, and I will continue my education at the University of California, Santa Barbara in the fall. If you asked me a week ago to name the parts of the eye I would have replied with “the pupil and the white part around it.” However, as my first week with the California Institute of Regenerative (CIRM) SPARK Program draws to a close, I now have a newfound understanding of the vast structures and knowledge related to the field of ophthalmology and ocular research. Thanks to the guidance I received from Dr. Adam Poe and the resources available at the Ghiam Lab at Cedars-Sinai I was able to discover the profound impact microRNA and LESCs (Limbal Epithelial Stem Cells) have on wound-healing in the epithelial layer of the Cornea. 

After reading some of the studies and research materials produced by the Ghiam Lab, the research and knowledge produced by the bright minds at Cedars-Sinai is truly remarkable and priceless. The highlight of my week was learning about how miRNA-146a has a crucial influence on the regulatory mechanisms of LESCs in wound-healing, especially in diabetic populations. It was amazing to see how the presence of a miRNA-146a inhibitor decreased the healing period by an average of 40% and probed my interest to delve further into the effects of microRNA on LESCs during the wound-healing process. In addition, the valuable insights provided by Dr. Tawackoli and Mr. Lawless on sterile techniques and the basics of stem cells will be vital throughout my time at the CIRM SPARK Program. I am ecstatic about joining the Cedars-Sinai team and cannot wait to continue learning about the role of LESCs in the epithelial layer of the cornea. 

Source: Winkler MA, Dib C, Ljubimov AV, Saghizadeh M. Targeting miR-146a to treat delayed wound healing in human diabetic organ-cultured corneas. PLoS One. 2014 Dec 9;9(12):e114692. doi: 10.1371/journal.pone.0114692. PMID: 25490205; PMCID: PMC4260878.

First Week; Hassan Samiullah

Hey there! I’m Hassan Samiullah, a rising 12^th Grader at Portola High School in Irvine, CA.

It took less than a day after the start of my internship at Cedars-Sinai through CIRM SPARK for me to be soaked in medical jargon. Indeed, my research under the mentorship of Dr. Katie Grausam of the Breunig Lab will focus on an intersection of neuroscience, cancer, stem cells, and molecular biology. While it is tempting to read over these words and dismiss them as being big words just for scientists (and/or nerds), I hope you can appreciate how they fit together like pieces of a puzzle.

Glioblastoma is a deadly type of cancer affecting glial cells—cells that support your brain’s neurons. Before testing any treatment against this cancer, scientists need tumor cells to work with. Enter GL261: a glioblastoma cell line originating from an experiment done over 80 years ago. In 1939, Dr. Arnold Seligman and Dr. M.J. Shear injected into mice a cariogenic substance called methylcholanthrene (try pronouncing that), sure enough causing brain tumors in 65% of the mice. Those tumor cells were preserved and have been used for research ever since. Cells from that very line glowed bright green on my laptop screen (as they were transfected by Dr. Grausam with mClover/GFP that inserted into the DNA) . I’ll get to see a mouse undergo brain surgery and be injected with those luminescent tumor cells (I’ll get the best seat in the house with Microsoft Teams), hopefully resulting in a tumor we can use for testing treatments. But just like all things in medicine (and in life), the mouse tumor model has limitations. Tumor cells from the GL261 line have mutations (changes in DNA that can lead to diseases like cancer) in the RAS and p53 genes. While p53 is a widely studied gene in cancer research, it turns out that RAS doesn’t commonly mutate in human glioblastomas: a study from 2013 found a mutation rate of just 1%.

Having mice tumor models that are like human tumors can make a study’s results more externally valid. Plus, it’s always good to have more data to work with. But is a different model possible? Enter plasmid DNA. It’s genetically engineered DNA that has mutations in genes similar to ones that can occur in human glioblastomas. I watched a laborious process of gene cloning to produce many of these plasmids, which are now being grown in bacteria, and which will hopefully be ready to give a mouse a tumor (this will involve electroporation: pretty much shocking cells so that plasmid DNA can pass through their membranes). I’m really excited to see how the two experiments will play out, namely how the GL261 tumor will compare with the plasmid DNA-induced tumor. Our overall goal is to see how the immune system interacts with tumors to affect not just the cancer itself, but also the environment around it, which can hopefully give us a better insight into how treatments can be more effective. Dr. Grausam has done a great job explaining her lab work and how it fits in to the bigger picture of fighting cancer, no matter how complicated the steps may be. I’m thankful for having her expertise, support, and feedback. I look forward to what we can learn and accomplish!

First Week; Aarav Dubey

Hello Everyone! My name is Aarav Dubey, and I have started my CIRM internship in the lab of Dr. Kumar under the mentorship of Dr. Shikhar Aggarwal at Cedars Sinai. The goal of my project is to understand and investigate immune responses and modulatory effects of macrophages during Acute Kidney Injury (AKI) repair/regenerative processes in experimental modals. Specifically, I will learn how macrophage-plasticity, spatiotemporally, impact the recovery processes after mammalian kidney injury. I am beginning to familiarize myself with different macrophage subtypes and how they can help or harm kidney recovery after injury. I am starting off this experience by reviewing and discussing various articles with my mentor to get a greater understanding of the cellular crosstalk in the injured milieu.  I have been having some difficulty understanding the language of some of these texts, but my mentor has helped me tremendously explaining the concepts. In the end, it feels very rewarding when I finally understood these concepts after spending many hours googling and discussing them with my mentor.