Dr. Rawlings’ primary research interests include dysregulated B cell development and signaling leading to immunodeficiency, autoimmunity or lymphoid malignancies, and the development of gene therapy for primary immune deficiency diseases. The Rawlings laboratory uses expertise in basic and clinical immunology, signal transduction and lymphocyte developmental biology to understand how altered signals can lead to immunologic disease, with the ultimate goal of developing translational therapies capable of specifically modulating these disorders. Dr. Rawlings is a member of multiple regional and national organizations as well as of an NIH study section, chairman for the USIDNET XLA patient registry, and ad-hoc reviewer for various grant programs and immunology journals. He also co-directs the Northwest Genome Engineering Consortium, a research program funded as part of the NIH Roadmap for Medical Research and comprised of seven collaborative projects focused on developing enzymatic reagents and delivery methods for site-specific gene repair in hematopoietic stem cells. Dr. David Rawlings pursues three major research themes: (1) B cell antigen receptor (BCR) engagement generates a multi-component complex of signaling effectors, or "signalosome." Current studies seek to understand better the components and interactions that constitute the signalosome through genetic and biochemical analysis of tyrosine kinases, adapter proteins, and lipid enzymes; and the use of animal models to evaluate the developmental consequences of altered expression of these proteins. A major focus of this current work is the multi-adapter protein CARMA1, which plays a crucial role in development of human B cell lymphomas. This work includes structural studies in collaboration with Roland Strong. (2) Human and murine in vitro B lineage culture models are used in conjunction with animal-based approaches (transgenic, knockout, and lentiviral vector-based RNA interference) to model B cell lineage development and B lymphoid malignancies. Current studies include analysis of thymic stromal lymphopoeitin-receptor (TSLP), Toll, Notch, and BAFF-receptor signaling cascades in the generation of peripheral B cell subsets and the PKC?/NF?B pathways in the development or progression of lymphoma. The work with TSLP involves collaboration with Dr. Steven Ziegler. (3) Gene therapy has the potential to correct primary immunodeficiency disorders. Because of the selective advantage for gene corrected cells in affected hosts, X-linked agammaglobulinemia and Wiskott-Aldrich Syndrome represent excellent disease targets for stem cell-based gene therapy. In addition to the use of cell lineage-specific viral vectors systems, we have begun to evaluate the potential of employing homing endonucleases to facilitate genetic repair of the mutant loci in animal models, in collaboration with Dr. Andrew Scharenberg and Dr. Nancy Maizels.
I) Representative publications on "Lymphocyte signal transduction" include:
Karen M. Sommer, Miguel E. Moreno- García, David J. Rawlings. The CARMA1 signalosome links the adaptive and innate signaling machinery in lymphocytes. (2006) Nature Reviews in Immunology (In Press)
Gomez TS, McCarney SD, Carrizosa E, Labno CM, Comiskey EO, Nolz JC, Zhu P, Freedman BD, Clark MR, Rawlings DJ, Billadeau DD, Burkhardt JK. HS1 Functions as an Essential Actin-Regulatory Adapter Protein at the Immune Synapse. (2006) Immunity 24:1-12.
Karen Sommer, Beichu Guo, Joel L. Pomerantz, Ashok D. Bandaranayake, Miguel E. Moreno-García, Yulia L. Ovechkina, David J. Rawlings. Phosphorylation of the CARMA1 Linker Controls NF-?B Activation. (2005) Immunity 23 (6): 561-574.
Humphries LA, Dangelmaier C, Kato RM, Griffith N, Irene Bakman I, Christoph W. Turk CW, Daniel JL, Rawlings DJ. Tec kinases mediate sustained calcium influx via site-specific tyrosine phosphorylation of the PLCg SH2-SH3 linker. (2004) J Biol Chem 279(36):37651-37661.
David J. Rawlings. The biology and biochemistry of inflammatory signolosomes. (2006) EMBO Reports 7 (1): 25-30. 6. Guo B, Su TT, Rawlings DJ. PKC family functions in B cell activation. (2004) Curr Opin Im
Saito K, Tolias KF, Saci A, Koon HB, Humphries LA, Scharenberg A, Rawlings DJ, Kinet J-P, Carpenter CL. Btk regulates PtdIns-4,5-P2 synthesis: importance for calcium signaling and PI3K activity. (2003) Immunity 19:669-678.
Su TT, Guo B, Chae K, Kawakami Y, Chae K, Kato RM, Kang SW, Patrone L, Wall R, Teitell MA, Leitges M, Kawakami T, Rawlings DJ. PKCb controls IkB kinase (IKK) lipid raft recruitment and activation in response to BCR signaling. (2002) Nature Immunol 3:780-786.
Kang, SW, Wahl, MI, Leitges M, Tarakhovsky, A, Tabuchi, R, Kato,R, Turck, CW, Kawakami, T, Witte ON, Rawlings, DJ PKCb modulates antigen receptor signaling via regulation of Btk membrane localization. (2001) EMBO 20:5692-5702.
Guo B, Kato RM, Garcia-Lloret M, Wall M and Rawlings DJ: Engagement of the human pre-B cell receptor (pre-B) generates a lipid raft-dependent calcium signaling complex. (2000) Immunity 13:243-253.
Rawlings DJ, Scharenberg AM, Park H, Wahl MI, Lin S, Kato RM, Fluckiger A-C, Witte ON, Kinet J-P: Activation of Btk by a phosphorylation mechanism initiated by src family kinases. (1996) Science 271:822-825. 12. Rawlings DJ, Saffran DC, Tsukada S, Largaespada DA, Grimaldi JC, Cohen L, Mohr RN, Bazan JF, Howard M, Copeland NG, Jenkins NA and Witte ON: Mutation of the BPK tyrosine kinase unique region in X-linked immunodeficiency. (1993) Science 261:358-361.
II) Representative publications on "Gene therapy for primary immunodeficiency disorders" include:
Miao, CH, Ye, P, Thompson, AR, Rawlings, DJ, Ochs, HD. Immunomodulation of transgene responses following naked DNA transfer of human factor VIII into hemophilia A mice. (2006) Blood 108(1):19-27.
Yu PW, Tabuchi RS, Kato RM, Astrakhan A, Chae K, Ellmeier W, Witte ON, Rawlings DJ. Sustained correction of B cell development and function in a murine model of X-linked agammaglobulinemia (XLA) using retroviral gene transfer. (2004) Blood 104(5):1281-1290.
Humblet-Baron S, Anover S, Kipp K, Zhu Q, Ye P, Zhang W, Ovechkina Y, Khim S, Astrakhan A, Strom T, Kohn D, Candotti F, Vyas Y, Ochs H, Miao C, Rawlings D. Lentiviral vector-mediated gene therapy as treatment for Wiskott-Aldrich Syndrome (WAS): Pre-clinical studies in human cell lines and WASp -/- mice. (2005) Molecular Therapy 11(1):S133.
Scharenberg AM, Rawlings DJ, Monnat RJ, Stoddard BL. Engineering and development of I-AniI homing endonucleases for gene correction applications. (2005) Molecular Therapy 11(1): S405.
Rawlings DJ. Bruton's tyrosine kinase controls a sustained calcium signal essential for B lineage development and function. (1999) Clinical Immunology 91:243-254.
III) Representative publications on "Modeling normal and altered lymphopoiesis" include:
Stephanie Humblet-Baron, Blythe Sather, Stephanie Anover, Shirly Becker-Herman, Debora J. Kasprowicz, Socheath Khim, Thuc Nguyen, Kelly Hudkins-Loya, Charles E. Alpers, Steve Ziegler, Hans Ochs, Troy Torgerson, Daniel J. Campbell, and David J. Rawlings. Wiskott Aldrich Syndrome Protein is Required for Regulatory T Cell Homeostasis. (2006) (Submitted)
Alexander Astrakhan, Miyuki Omori, Thuc Nguyen, Shirly Becker-Herman, Masanori Iseki, Theingi Aye, Kelly Hudkins-Loya, James Dooley, Andrew Farr, Charles E. Alpers, Steve Ziegler, and David J. Rawlings. Local Elevation in Thymic Stromal Lymphopoietin Induces Systemic Alterations in B Cell Development. (2006) (Submitted)
Su TT, Guo B, Wei B, Braun JB, Rawlings DJ. Signaling of transitional type 2 B cells is critical for peripheral B cell development. (2004) Immun Rev 197:167-178.
Hoyer KK, French SW, Turner DE, Nguyen MTN, Renard M, Malone CS, Knoetig S, Qi CF. Su TT, Cheroutre H, Wall R, Rawlings DJ, Morse HC, and Teitell MA. Dysregulated TCL1 Promotes multiple classes of mature B cell lymphoma. (2002) Proc Nat Acad Sci USA 99: 14392-14397.
Su TT, Rawlings DJ. Transitional B lymphocyte subsets operate as distinct checkpoints in murine splenic B cell development. (2002) J. Immunol. 168: 2101-2110.