Heart disease is the single largest killer of Americans. Those that survive a heart attack have a higher risk of another attack heart failure, and stroke. Consequently, there is a need to develop approaches for repairing the heart and/improving the quality of life after a heart attack. The concept of regenerative medicine through the use of stem cells is gaining attention as a potential therapy for restoring cardiac function. Hurdles currently faced in the development of cellular therapies for the clinic are (1) the limited availability of cell surface markers for selecting pure cell populations and for tracking differentiation and (2) the lack of knowledge of how in vitro differentiation and in vivo development of cardiomyocytes correlate. The long term goal of this project is to correlate the function of a pure population of stem cells with a panel of defined cell surface markers. The major objective of the first granting period (K99) is to address the need for informative cell surface markers that are useful for characterizing specific cell types, stages, and lineages. I will apply the experience and knowledge gained from the research in the first granting period to understanding stem cell development in the second phase (ROO).
The aims of the ROO phase are designed to develop an understanding of whether the molecular events during in vitro differentiation mimic those during in vivo development of cardiomyocytes. My long term career goal is to lead an independent research group that utilizes analytical technologies to bridge basic science research and clinical applications of stem cells. This research plan is specifically designed to take advantage of my analytical expertise while providing a relevant biological context to acquire new knowledge and skills necessary maximize the use of analytical methods within the context of biology. The training environment at Johns Hopkins is especially suited to this research due to the access to state-of-the art instrumentation, mentors with specialized expertise, and the forum to support discussion of clinically relevant research. Relevance: The focus of the current proposal is to identify specific cell surface protein markers that will aid clinicians and researchers in understanding the mechanisms of stem cell differentiation, both within the animal and in culture. Accessible markers for the specific selection, enrichment, and tracking of stem cell populations appropriate for cardiovascular regenerative medicine will be generated.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Career Transition Award (K99)
Project #
5K99HL094708-02
Application #
7759516
Study Section
Special Emphasis Panel (ZHL1-CSR-Z (O1))
Program Officer
Commarato, Michael
Project Start
2009-01-21
Project End
2010-12-31
Budget Start
2010-01-01
Budget End
2010-12-31
Support Year
2
Fiscal Year
2010
Total Cost
$98,004
Indirect Cost
Name
Johns Hopkins University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Boheler, Kenneth R; Gundry, Rebekah L (2017) Concise Review: Cell Surface N-Linked Glycoproteins as Potential Stem Cell Markers and Drug Targets. Stem Cells Transl Med 6:131-138
Mallanna, Sunil K; Waas, Matthew; Duncan, Stephen A et al. (2017) N-glycoprotein surfaceome of human induced pluripotent stem cell derived hepatic endoderm. Proteomics 17:
Mallanna, Sunil K; Cayo, Max A; Twaroski, Kirk et al. (2016) Mapping the Cell-Surface N-Glycoproteome of Human Hepatocytes Reveals Markers for Selecting a Homogeneous Population of iPSC-Derived Hepatocytes. Stem Cell Reports 7:543-556
Gundry, Rebekah L; Tchernyshyov, Irina; Sheng, Shijun et al. (2010) Expanding the mouse embryonic stem cell proteome: combining three proteomic approaches. Proteomics 10:2728-32
Gundry, Rebekah L; Raginski, Kimberly; Tarasova, Yelena et al. (2009) The mouse C2C12 myoblast cell surface N-linked glycoproteome: identification, glycosite occupancy, and membrane orientation. Mol Cell Proteomics 8:2555-69
Gundry, Rebekah L; White, Melanie Y; Murray, Christopher I et al. (2009) Preparation of proteins and peptides for mass spectrometry analysis in a bottom-up proteomics workflow. Curr Protoc Mol Biol Chapter 10:Unit10.25