BD Biosciences Research Grants

Summer 2015 Research Grant Recipients Talk About Their Research


Shangqin Guo, PhD
Assistant Professor, Department of Cell Biology and Yale Stem Cell Center
Yale University

Abstract Title:
Identifying Cell Surface Markers for the Purification of Privileged Cells


BD: What is your educational background?

Shangqin Guo: I graduated from Sichuan University, Chengdu, China, with a bachelor of science degree in biology. After that, I pursued my PhD training at the Department of Biochemistry, Boston University, in Massachusetts. After my doctoral training, I ventured into the field of stem cell biology through a postdoctoral position in the lab of Dr. David Scadden at the Center for Regenerative Medicine, Massachusetts General Hospital and Harvard Stem Cell Institute.

BD: How and when did you become interested in science?

Shangqin Guo: I can’t think of any special person, event, or class. My interest in science just came naturally. Maybe my family had some influence, as my grandfather was a very well respected doctor and I felt drawn toward understanding how medicine works in a more fundamental way.

BD: How did you become interested in your broad field of study?

Shangqin Guo: I remember being intrigued with the question what is life and how does it perpetuate itself? In mammalian cells, I knew cancers and stem cells could do that. In some sense, I wanted to know what was giving cancers and stem cells the property of self-perpetuation. Having completed my PhD training in the field of cancer biology, where everything is supposed to be quite messed up, I thought I’d try out stem cell biology to gain a perspective of how normal cells manage to self perpetuate. However, my appreciation for biology at the time was very vague and I could not articulate a clearly defined and tangible question to go after. Shortly after I began my postdoctoral training in stem cell biology, the landmark discovery of pluripotency induction by defined transcription factors was reported by Dr. Shinya Yamanaka and colleagues. I listened to a lecture given by Dr. Yamanaka at Mass General shortly after his study appeared. I think I’m just one of many scientists that were immediately taken by the simplicity, power and profound implication of this breakthrough.

BD: Describe the project for which you received the BD grant.

Shangqin Guo: In 2006, Dr. Shinya Yamanaka reported the landmark discovery that fibroblasts can be turned into pluripotent stem cells. Even though this amazing change can occur, it takes a long time and very few cells can manage the change. We wanted to know what is special about the cells that “forget” their somatic identity and switch to pluripotency. With a new imaging system, we were able to watch the full transformation process – how a blood cell changed into a pluripotent stem cell – step by step. We learned that rare somatic cells can undergo this change rather efficiently. Why would these cells be so willing to forget they are blood cells and readily change into something that is very different? Do such cells only exist in the blood tissue, or they are also present in other tissue types? We hope in this project to identify a panel of cell surface markers that will allow us to identify which cells readily switch their identities and to purify these cells for further study of their unique biology and the Yamanaka reprogramming process.

BD: What are the long- and short-term scientific goals of this project?

Shangqin Guo: The short-term goal is to identify a panel of cell surface markers and validate their value in purifying the relevant cell populations. The long-term goal is to understand the molecular details of the very few cells that change their identities. Ultimately, we hope to understand the general rules of how a cell decides to maintain its original destiny or change into something else.

BD: What are the implications of your project for human health?

Shangqin Guo: First, understanding how somatic cells can change their identity to pluripotent stem cells would improve our ability to isolate these cells for research and potential therapies. Second, the pluripotency induction studies provide the opportunity to understand the more general rules of how cell types interconvert. If these rules are sufficiently understood, any cell type should in principle have the ability to convert into any other cell type. For example, we specifically use blood progenitor cells obtained from bone marrow to derive pluripotent stem cells. If we know the rules sufficiently well, we would like to derive from these blood progenitor cells various cell types that are damaged by disease or injury, such as pancreatic beta cells, neurons, or cardiomyocytes. Third, we might learn how to prevent the emergence of unwanted cell types, such as malignant cells or overly inflammatory cells.

BD: Which BD reagents do you plan to use, and for what purposes?

Shangqin Guo: We will use a wide variety of antibodies, buffers and other reagents for flow cytometry analysis and sorting to characterize the cell surface phenotypes of the unique somatic cells that do change their identity. After successful validation, we will also use these markers to prospectively isolate or enrich for these remarkable cells to be used for the derivation of therapeutic cell types, as well as for studying the molecular details about how cells change.


Ae-Ri Ji, PhD
Postdoctoral Researcher
University of Granada Junta de Andalucia

Abstract Title:
Human iPSC-based Model to Develop a Personalized Treatment for Ménière's Disease by Sustained Target Gene Knockdown


BD: What is your educational background?

Ae-Ri Ji: I received my undergraduate degree in chemistry in 2000 from Dankook University in Korea, my master’s degree in biochemistry in 2003 from Yonsei University, and my Ph.D. in human embryonic stem cell biology from Seoul National University in 2012. After spending some time as a research professor at Dankook U., I moved to the Pfizer Center at the University of Granada Junta de Andalucia Centre for Genomics and Oncological Research (GENYO) as a postdoctoral researcher. I currently work under the direction of Dr. Jose A. Lopez-Escamez. My main field of study is regeneration of the central nervous system using human pluripotent stem cells (hPSCs).

BD: How and when did you become interested in science?

Ae-Ri Ji: I made the decision during a difficult time in my life, between 2006 and 2007. Thanks to my work in science I was able to overcome these hard times. From that point onward, I believed that being a scientist was my destiny.

BD: How did you become interested in your broad field of study?

Ae-Ri Ji: When I began studying human embryonic stem cells (hESCs), I saw the cardiac myocyte which was differentiated from hESCs and beating in the dishes. At that moment, my heart was beating, too. I'll never forget that memory. After finishing my doctoral studies, I developed even stronger feelings about the importance of stem cell research. I decided that I would use stem cells as tools for studying specific diseases, particularly for regenerative medicine. That is what I am doing in this postdoctoral position.

BD: Describe the project for which you received the BD grant.

Ae-Ri Ji: Meniere’s disease (MD) is a chronic illness characterized by episodes of vertigo lasting from minutes to hours, with fluctuating sensorineural hearing loss, tinnitus, and aural pressure. When symptoms are severe they usually interfere with normal life functioning. There is no effective treatment for MD. Histopathological studies in human temporal bones have demonstrated an accumulation of endolymph in the cochlear duct, leading to fluid accumulation in the cochlea and the vestibular end organs. The underlying mechanisms for fluid buildup, known as hydrops, remain unknown.

Most cases of MD are sporadic, meaning they occur in individuals with no family history of the disease. But a small percentage of all cases are known to run in families. When the disorder is familial (FMD) it most often has an autosomal dominant pattern of inheritance. Autosomal dominant inheritance means one copy of an altered gene in each cell is sufficient to increase the risk of the disorder. However, until recently no associated genes have been identified. Using exon sequencing my team has discovered four novel candidate genes in three families whose members suffer from MD, and validated them through Sanger sequencing. However, the exact pathogenesis of these missense variants in MD has not been characterized.

Many MD studies employ mouse or guinea pig animal models, which have the advantage of providing an in vivo disease model. However, animal models show significant differences from humans, for example, as in this disease, the exact duplication of the mutation of a target gene. Therefore animal models are probably not appropriate for investigating FMD. Similarly primary cultured cells from animal or human cochlea tissue are heterogeneous populations and are difficult to obtain in quantities sufficient for studying MD. Therefore, the development of human cell model in vitro is required to investigate MD disease characteristics and to understand the mechanism of FMD caused by specific target gene mutations. Furthermore, patient-specific cell models can overcome the limitation of animal models and primary cultured cells.

Induced pluripotent stem cells (iPSCs) are self-renewable and their pluripotency allows them to differentiate into specific cell types in sufficient numbers for subsequent studies. Human iPSCs can be obtained directly from somatic cells by introducing defined transcription factors Oct4, Sox2, Klf4, and c-Myc. Hence, the generation of human iPSCs from somatic cells offers a powerful system for investigating human genetic diseases in vitro.

BD: What are the long- and short-term scientific goals of this project?

Ae-Ri Ji: Our short-term goals include generation of FMD patient-specific-iPSC lines with one or two target gene mutations. We also hope to characterize the differences between FMD-iPSC and control-iPSC lines. Long term, we hope to differentiate FDM- and control-iPSCs into epithelial otic progenitor cells and neural otic progenitor cells. We also expect to identify genetic and phenotypic differences, and uncover the molecular mechanism through which mutated genes cause FMD. I expect that a knock-in of a wild-type gene into patient-iPSC lines using the CRISPR/Cas9 system will be able to restore the phenotype.

BD: What are the implications of your project for human health?

Ae-Ri Ji: These studies will show that several mechanisms are responsible for generating the MD phenotype. Eventually, we hope that iPSC cell models for FMD will be used in the development of new drugs and perhaps personalized therapies for MD.

BD: Which BD reagents do you plan to use, and for what purposes?

Ae-Ri Ji: For feeder-free culture of established iPSC lines and to induce differentiations into epithelial otic progenitors, neural otic progenitors (NOP), inner ear hair cells we plan to use Matrigel® matrix (Corning), and various antibodies to identify and characterize iPSCs or iPSC-induced precursor cells SSEA-4, Tra-1-60, CD30, Pax8, Sox2, ATOH1 and BRN3C, BRN3C and MYO7A, BRN3A and NeuroD1. We will also use BD instrumentation such as the BD FACSCanto™ II flow cytometer, BD FACSVerse™ flow cytometer, and BD FACSAria™ cell sorter to characterize and isolate live cells with high viability.

Matrigel® is a registered trademark of Corning Incorporated

Evgenia Verovskaya, PhD
Postdoctoral Fellow
The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco

Abstract Title:
Contribution of the Bone Marrow Niche to Hematopoietic Stem Cell Aging


BD: What is your educational background?

Evgenia Verovskaya: I received a degree in Pharmacy from the Sechenov First Moscow State Medical University, Moscow, Russia, in 2006. For my graduate studies I moved to the Netherlands, to the University of Groningen, where I received an MSc in 2008 in medical and pharmaceutical drug innovation and in 2014 a PhD in medical science. My doctoral project, in the laboratory of Prof. Gerald de Haan, addressed the clonal contributions of young and old hematopoietic stem cells (HSCs) to blood formation using a barcoding technique. In 2014, I joined the laboratory of Prof. Emmanuelle Passegué at the University of California, San Francisco, as a postdoctoral fellow. My postdoctoral project focuses on understanding the interplay between old HSCs and their bone marrow microenvironment, and the contribution of that interaction to blood aging.

BD: How and when did you become interested in science?

Evgenia Verovskaya: With my mother being a physician and my father a scientist, I was exposed to scientific questions at a very early age and became quickly fascinated by what drives human diseases. After many years studying science, I still find the questions relevant to human health the most interesting to study.

BD: How did you become interested in your broad field of study?

Evgenia Verovskaya: My interest in stem cell biology began during my graduate studies at Groningen. I started this program the year Prof. Shinya Yamanaka discovered induced pluripotent stem cells (iPSCs), for which he received a Noble Prize. My interest in this topic lead me to an internship in the laboratory of Prof. Gerald de Haan, where I became experienced with blood-forming stem cells. During this internship, I became fascinated by the idea that a single HSC could repopulate the entire blood system of an irradiated mouse. I then decided to join Prof. de Haan’s laboratory for my PhD work to continue studying HSCs, in particular their clonal heterogeneity. My project involved quantifying differences in repopulation behavior between HSCs isolated from young and old mice using DNA barcodes. Using this method, we found profound heterogeneity in differentiation potential and repopulation capacity between individual HSCs. We also documented differences in HSC pool size, and migratory behavior of young and old HSCs.

BD: Describe the project for which you received the BD grant.

Evgenia Verovskaya: Aging is manifested in the blood system by an increased rate of anemia, blood cancers, autoimmune diseases, and impaired ability to fight infection. Such declines have been linked to the diminished function of aged HSCs. Understanding the molecular mechanisms and factors driving HSC functional decline is essential to developing strategies to mitigate blood aging, in particular to prevent blood cancers and restore the ability to fight infections in the elderly. Recent studies have shown that the bone marrow (BM) microenvironment in which HSCs reside, which is also known as the HSC niche, is essential for maintaining HSC function and controlling blood production. BM niche cells provide important instructive cues to the HSCs that regulate the size of the HSC population, their cycling activity, and their differentiation behavior.

Pathological remodeling of the BM niche can lead to HSC exhaustion or the development of blood cancers or leukemia. Recently, our laboratory identified important crosstalk mechanisms between the leukemic BM niche and HSCs, which directly contribute to disease development and the loss of normal HSC function. In this project, I am investigating whether similar cross-talk mechanisms exist between the aging BM niche and HSCs.

I am testing the hypothesis that physiological aging drives the remodeling of the HSC niche, both in terms of cellular components and secreted/expressed regulatory factors, which, in turn, drive or reinforce HSC aging features thereby directly contributing to blood aging. To address these questions, I am investigating different populations of BM niche cells for changes in numbers, function, and molecular regulation. I am also directly testing whether targeting the changes in the aging BM niche will help rejuvenate old HSCs and improve blood aging.

BD: What are the long- and short-term scientific goals of this project?

Evgenia Verovskaya: My immediate goal is to understand at the global level how changes in the BM microenvironment contribute to HSC aging and to identify the molecular mechanisms involved. My long-term goal is to establish whether targeting the aging BM niche can be used to prevent or reverse blood aging and rejuvenate aged HSCs.

BD: What are the implications of your project for human health?

Evgenia Verovskaya: Aging populations are a hallmark of the twenty-first century. Aging is associated with decreased regeneration and increased incidence of a range of debilitating and chronic diseases that impair quality of life and strain economic and medical resources. Preventing or reversing age-related defects therefore holds tremendous potential therapeutic benefit. One emerging characteristic of aging is that reduction in tissue function usually correlates with a reduction in stem cell activity, resulting in diminishment of tissue integrity and function leading to physiological decline. Several recent studies have shown that restoration of youthful levels of signaling in aged stem cells in muscle and brain can partially reverse their aging features. This project will contribute to our understanding of the mechanisms of stem cell aging focusing on the blood system and its rare population of blood-forming HSCs, and may open the way to develop new therapies for preventing or reversing blood aging through manipulation of the bone BM microenvironment.

BD: Which BD reagents do you plan to use, and for what purposes?

Evgenia Verovskaya: I expect to use a whole range of fluorophore-conjugated BD antibodies to isolate, image and functionally investigate HSCs and their BM niche cells. I will also use BD™ Cytometric Bead Array and BD OptEIA™ ELISA kits to investigate changes in pro-inflammatory cytokines in the aging BM niche.


Huan (Sharon) Wang, PhD
Postdoctoral Research Fellow
Harvard University Medical School

Abstract Title:
Single Cell Network Modeling of Drug-Induced Cardiotoxicity


BD: What is your educational background?

Huan Wang: I received my BS in biotechnology from Zhejiang University in Hangzhou, China. During this time, I also spent one year as an exchange student in the Chinese University of Hong Kong, where I was exposed to different cultures and education styles. My college studies equipped me with fundamental knowledge not only in biology, but also in chemistry, math and physics. I left China in 2006 for the University of Colorado, Boulder, where I earned my PhD in molecular, cellular and developmental biology in 2013. My doctoral thesis focused on understanding cardiac valve calcification using experimental techniques in both molecular biology and biomaterial engineering. I am currently a postdoc working under the mentorship of Dr. Peter Sorger in the Laboratory of Systems Pharmacology. Our main goals are to determine how cancer drugs cause unwanted cardiotoxicity and how we can develop therapeutic alternatives by better understanding the mechanisms of toxicity.

BD: How and when did you become interested in science?

Huan Wang: I became interested in science in high school, where I found that biology class was fun. I had a very dedicated and patient teacher who related many interesting stories about biological discoveries. I also gained a lot of confidence as I was able to answer most of the questions in class. After finishing the stressful college entrance exam that millions of Chinese students take every year, I chose biology as my major. Thus began my journey in the biological sciences.

BD: How did you become interested in your broad field of study?

Huan Wang: In 2012, Drs. Gurdon and Yamanaka shared the Nobel Prize in Physiology or Medicine for their discovery that mature human somatic cells, eg, fibroblasts, can be transformed into induced pluripotent stem cells (iPSCs). This technique is revolutionizing medicine by creating superior platforms for drug screening, tissue regeneration, and biological research. For example, patient-derived iPSCs differentiate into different human tissue cell types and are used for toxicity screening in vitro. In addition, iPSCs could accelerate multiple tissue repair and regeneration after injury or under certain stressful conditions. Because of this great medical potential, I became interested in stem cell research and more specifically, its applications in cardiovascular repair. Heart complications are still the number one cause of human death in the US. Besides genetic causes, cancer drug treatment is another leading cause of cardiac damage and deterioration of cardiac output. Understanding the underlying molecular mechanisms of how and why cancer drug treatment leads to cardiovascular toxicity can bring new insight on the clinical treatment for these complications.

BD: Describe the project for which you received the BD grant.

Huan Wang: Potent cancer drugs come at the price of unexpected toxicity, particularly to the heart. Many cancer patients are not seen by a cardiologist early enough to avoid or detect cardiovascular issues resulting from taking cancer drugs. This is primarily due to the lack of understanding on how and why cancer drugs lead to cardiotoxicity. We are using human stem cell-derived cardiomyocytes, which closely mimic the native physiology of cardiac cells, to study the underlying molecular targets and mechanisms. We are measuring changes in signaling and gene expression in response to drug perturbations in these cells, which could help us identify signatures of changes that could serve as major causes in cardiotoxicity. One hypothesis that we want to test is whether the drugs cause cardiac damage through the same mechanisms by which they kill cancer cells. When drugs reach the heart they elicit cascades of changes in signaling proteins and gene expression. We plan to use antibodies provided by BD to measure and quantify those signal changes in both cultured cells and patient biopsies. By probing these biological systems broadly and in an unbiased manner, we hope to learn the causes of drug-induced cardiotoxicity.

BD: What are the long- and short-term scientific goals of this project?

Huan Wang: The long-term goal of the project is to develop strategies or medical treatment that can mitigate cardiotoxic effects of cancer drugs. In the short-term, we are working closely with computational scientists and clinical researchers to fully understand our experimental data and build biological models that explain the molecular mechanisms of cardiotoxicity. In the meantime, through drug perturbations, we may also come to understand the basic biology of how cardiac cells function normally and under stressed conditions.

BD: What are the implications of your project for human health?

Huan Wang: Based on current clinical data, cardiovascular-related mortality is seven-fold higher in age-matched pediatric cancer patients than in normal individuals. In addition, approximately twelve percent of breast cancer survivors suffer heart failure within three years of chemotherapy treatment. The implications of this project are threefold. First we hope to help patients undergoing cancer therapy to survive the potentially lethal heart problems. We will study the molecular mechanisms of drug-induced cardiotoxicity and propose and test novel treatment regimes that may reduce or prevent cardiotoxicity. Next, with the cardiotoxicity biomarkers that we identify, we could improve the process of drug development and select for safer, less toxic drugs. Finally, our discoveries in the domain of cardiovascular toxicity induced by cancer drugs may be relevant or even shared by toxicity observed in other tissues, such as kidney and liver.

BD: Which BD reagents do you plan to use, and for what purposes?

Huan Wang: We plan to use the primary antibodies from BD to examine signaling pathway changes in response to drug treatment based on flow cytometry or immunofluorescence. We believe that we can reveal how cardiac systems function under the stressful condition of cancer drug treatment through the proposed experiments.


Xiao Yan, PhD
Visiting Graduate Researcher
University of California, Los Angeles

Abstract Title:
Growth Factor Receptor-bound Protein 10 (Grb10) Regulates Hematopoietic Stem Cell Renewal


BD: What is your educational background?

Xiao Yan: I received my undergraduate degree in biological sciences from Peking University, Beijing, China, in 2010. I then moved to the US where I earned my PhD in pharmacology at Duke University in 2015 under the tutelage of Prof. John Chute. My doctoral thesis work involved hematopoietic stem cells. I am currently a visiting graduate researcher at UCLA, where I continue studying hematopoietic stem cells with Dr. Chute.

BD: How and when did you become interested in science?

Xiao Yan: During college, I was at first unsure of my career path, so I tried several things. I sold newspapers, worked in different labs, and took several random classes to see what I was truly interested in. During my senior year, I went to Stowers Institute, Kansas City for an internship, where I studied novel signaling pathways controlling protein degradation. Stowers was sponsored and built by billionaires James and Virginia Stowers, both cancer survivors, who donated almost all their fortune to build a state-of-the-art research facility. Through their generosity, I realized the significance of scientific research, especially biomedical research, and decided to make a career out of fighting serious diseases.

BD: How did you become interested in your broad field of study?

Xiao Yan: After my admission to the Duke Pharmacology program, I did a few rotations during which I worked in different settings ranging from basic biochemical labs to preclinical research labs. After the first year, I gradually realized that I was mostly excited about the translation of basic research to clinical applications. I understood that basic research is fundamental, but I was more eager to work on something that is closely related to drug development. At that time interest in cellular therapy was rising and I thought it was a promising field to enter. Traditional medicine, which focuses on small or big molecules, is too limited because of the complexity of biological systems. I believe a therapeutic method on the cellular level or organ level was the next logical step. Bone marrow transplantation has been the most mature cellular therapy method thus far, which is why I choose to work on hematopoietic stem cell research.

BD: Describe the project for which you received the BD grant.

Xiao Yan: A few years ago a research group at Baylor University proposed the idea that a family of imprinted genes, which are only expressed from the allele inherited from mother or father, are the key to determine the “stemness” of stem cells. Since then, no research has supported this theory. To attack this problem I plan to focus on one promising imprinted gene, Grb10. I hypothesize that Grb10 is a key regulator for stem cells. Using virus-mediated knock down of the Grb10 expression, I have shown that Grb10 deficiency leads to the depletion of hematopoietic stem cells. With a mouse model in which Grb10 is knocked out, I observed the same phenomenon: Grb10 disruption leads to a decrease of stem cells in the bone marrow. To my surprise, Grb10-deficient mice are strongly protected against radiation. After a radiation dose that kills most of the hematopoietic stem cells, mice lacking Grb10 recover their hematopoietic stem cells faster than the control mice – a discovery that may lead to new therapeutics.

BD: What are the long- and short-term scientific goals of this project?

Xiao Yan: The long-term goal for my project is to develop new therapeutic methods for radiation treatment. From the preliminary data I have at this moment, the most possible way is to screen inhibitors that bind to Grb10. As for the short-term goal, we want to determine the mechanism through which Grb10 regulates hematopoietic stem cells. We believe this could help us decipher the relationship between gene imprinting and stemness.

BD: What are the implications of your project for human health?

Xiao Yan: There are two main implications of my project. The successful rate of bone marrow transplantation is limited by the low populations of hematopoietic stem cells. Therefore amplification of stem cell populations has long been a research goal. Based on our work, Grb10 is a target that can be manipulated to regulate hematopoietic stem cell proliferation. For example, hematopoietic stem cells cannot be amplified in cell culture because they tend to differentiate, losing their stemness. However, if we could manipulate the expression level of Grb10, perhaps we could produce more stem cells in culture while maintaining their stemness.

Nowadays, the threat of nuclear war has become a real issue. A quick and efficient treatment plan for people suffering from radiation exposure is needed. We believe Grb10 is important for irradiation recovery and that inhibiting Grb10 after irradiation could be a powerful way to promote stem cell recovery, thus increasing the survival rate.

BD: Which BD reagents do you plan to use, and for what purposes?

Xiao Yan: We will perform KSL and SLAM KSL analyses to examine the hematopoietic stem cells. The assays will be performed as follows: bone marrow cells are incubated with BD Pharmingen™ and BD Horizon™ antibodies anti-c-kit, anti-sca-1, anti-lineage, anti-CD48, and anti-CD150. Peripheral blood samples from bone marrow transplantation assays will be incubated with BD Pharmingen™ antibodies anti-CD45.2, anti-CD45.1, anti-CD3, anti-Mac1, anti-Gr1, anti-Ter119 and anti-B220. This allows us to determine engraftment level. Cell cycle and cell death analyses will be conducted with BD Pharmingen Ki-67 set and AnnexinV: Apoptosis Detection kit I. In the phosphorylation assays, BD Phosflow™ anti-pAkt, anti-pS6 and anti-p4EBP1 will be used to detect the mTOR pathway activity. Data will be collected by BD FACSCanto™ II cell analyzer and analyzed by BD FACSDiva™ software version 8. HSC enrichment is achieved by running the cells through BD FACSAria™ II cell sorter.