The long term goal of this proposal is to train the applicant to become an independent academic physician- scientist studying the innate immune system and its impact on blood cancers. The principal investigator (PI) has completed PhD training focused on the cellular biology of natural killer (NK) cells, and MD training in internal medicine and hematology-oncology. This application describes a 5 year training program that will provide a mentored educational experience aimed at developing new scientific expertise in molecular profiling, massively parallel sequencing, proteomic analysis, manipulation of microRNAs (miRs), and the generation of mouse models. Dr. Timothy Ley will mentor the Pi's scientific and career development. He is a recognized leader in the field of lymphocyte cytotoxicity, in vivo genetic mouse models, and genomic analysis of leukemia. Furthermore, an advisory committee of medical scientist experts will provide additional scientific and and non-scientific career development guidance. The proposed research will evaluate the role of miRs in the regulation of NK cell cytokine activation. Recent work by the PI in Dr. Ley's laboratory identified two critical cytotoxic molecules, granzyme B (GzmB) and perforin (Prf1), that are post-transcriptionally regulated in NK cells. We hypothesize that miRs regulate GzmB and Prf1 in resting NK cells, and that cytokine-activation releases their block in translation. To address this hypothesis, we propose the following specific aims: 1) We will define the miR expression profiles in resting and cytokine-activated NK cells, and evaluate candidate miRs that may regulate GzmB and Prfl mRNA translation. 2) We will define the mRNA (transcriptome) and protein (proteome) expression profiles of resting and cytokine-activated NK cells, integrate these databases to define the mode of regulation of molecules important during NK cell activation, and define the role of miRs for post-transcriptional regulation. Techniques utilized in the project include miR sequencing, miR microarrays, in vitro and in vivo manipulation of miRs, the generation of a NK cell-specific Cre mouse model, the generation of genetic mouse models deficient in miRs using Cre-Lox, and the global analysis of the NK cell transcriptome and proteome. Washington University provides an ideal setting to train physician-scientists, and will foster an invaluable mentored eduational experience for the PI to realize his career goals in academic medicine.

Public Health Relevance

This overall career development proposal will train an independent physician-scientist for a lifetime of research studying the immune system and cancer. As NK cells are key components of immunity to numerous infectious pathogens, and are involved in the immunosurveillance of malignancy, the research proposed may have far reaching consequences for health and disease. Specifically, a better understanding of NK cell activation may lead to novel immune based stategies to treat hematologic malignancies.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08HL093299-04
Application #
8244451
Study Section
Special Emphasis Panel (ZHL1-CSR-O (F1))
Program Officer
Welniak, Lisbeth A
Project Start
2009-04-26
Project End
2014-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
4
Fiscal Year
2012
Total Cost
$116,503
Indirect Cost
$8,630
Name
Washington University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Leong, Jeffrey W; Chase, Julie M; Romee, Rizwan et al. (2014) Preactivation with IL-12, IL-15, and IL-18 induces CD25 and a functional high-affinity IL-2 receptor on human cytokine-induced memory-like natural killer cells. Biol Blood Marrow Transplant 20:463-73
Leong, Jeffrey W; Sullivan, Ryan P; Fehniger, Todd A (2014) microRNA management of NK-cell developmental and functional programs. Eur J Immunol 44:2862-8
Sullivan, Ryan P; Fogel, Leslie A; Leong, Jeffrey W et al. (2013) MicroRNA-155 tunes both the threshold and extent of NK cell activation via targeting of multiple signaling pathways. J Immunol 191:5904-13
Sullivan, Ryan P; Leong, Jeffrey W; Schneider, Stephanie E et al. (2012) MicroRNA-deficient NK cells exhibit decreased survival but enhanced function. J Immunol 188:3019-30
Fehniger, Todd A; Larson, Sarah; Trinkaus, Kathryn et al. (2011) A phase 2 multicenter study of lenalidomide in relapsed or refractory classical Hodgkin lymphoma. Blood 118:5119-25
Fehniger, Todd A; Wylie, Todd; Germino, Elizabeth et al. (2010) Next-generation sequencing identifies the natural killer cell microRNA transcriptome. Genome Res 20:1590-604
White, Douglas W; Keppel, Catherine R; Schneider, Stephanie E et al. (2010) Latent herpesvirus infection arms NK cells. Blood 115:4377-83