This K01 application is designed to prepare the applicant with the skills necessary to establish an independent research program on the study of systemic lupus erythematosus (SLE). SLE is an autoimmune disease of unknown cause which mainly afflicts women in their child bearing years and affects multiple organ systems of the body. T cell dysfunction due to abnormally altered intracellular signaling events is thought to be central in the pathogenesis of this disease. A critical signaling molecule known as the CD3 zeta chain is expressed in abnormally low amounts in the T cells of SLE patients, and this defect contributes to the poor functional capacity of the SLE T cells. Expression of an abnormal alternatively spliced (AS) form of the CD3 zeta mRNA with poor half life is a contributor to the reduced expression of zeta chain in SLE T cells. Aberrant splicing is a common feature observed in SLE and involves other critical genes involved in T cell signaling, migration, and cytokine production and the regulation of these genes has not been characterized. To understand the regulation of the CD3 zeta alternative splicing, the applicant used a discovery approach and identified several proteins binding to the CD3 zeta mRNA. Among these proteins, a promising candidate is the serine arginine (SR) family member Alternative Splicing Factor/Splicing Factor 2 (ASF/SF2). The applicant published that in T cells from healthy individuals, ASF/SF2 regulates CD3 zeta chain expression by suppressing production of the AS isoform. Examination of T cells from SLE patients revealed that patients had reduced expression of ASF/SF2 suggesting that aberrant ASF/SF2 expression may contribute to their defective T cell function. Additionally, the ASF/SF2 expression levels appeared to inversely correlate with the disease activity of SLE patients. The applicant's preliminary studies have shown a novel role for the splicing regulator ASF/SF2 in T cell physiology with a potential role in SLE pathophysiology. Based on the preliminary evidence, the hypothesis of this project is that ASF/SF2 regulates human T cell function and may represent a contributor to the SLE T cell defect. Toward the hypothesis, this project is aimed to understand the role of ASF/SF2 in human T lymphocytes and to determine whether ASF/SF2 may represent a disease marker in SLE. Methods will include biochemical analyses of human T cells and the development of a cell line to enable controlled expression of ASF/SF2.
The specific aims of the project are 1) To determine whether TCR stimulation regulates ASF/SF2 expression and activity in human T lymphocytes 2) To determine how ASF/SF2 regulates T cell function and 3) To determine whether ASF/SF2 expression levels in SLE T cells may represent a disease marker. To accomplish these aims, the applicant will need to expand her technical and intellectual skills in cellular and molecular immunology and become proficient in the design, interpretation and analysis of studies in SLE patients. The research environment at Beth Israel Deaconess Medical Center and the other affiliates of the Harvard Medical School system will provide the applicant with the resources to reach her goals within 5 years.

Public Health Relevance

Systemic lupus erythematosus is a chronic disease affecting over 1 million Americans. It afflicts mainly women in their child-bearing years and causes painful arthritis, with potentially fatal complications in the kidneys. The proposed project seeks to understand the potential role of an important protein ASF/SF2 in the T cells of SLE patients to assess its usefulness in disease management.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01AR060781-02
Application #
8462912
Study Section
Arthritis and Musculoskeletal and Skin Diseases Special Grants Review Committee (AMS)
Program Officer
Mancini, Marie
Project Start
2012-05-01
Project End
2017-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
2
Fiscal Year
2013
Total Cost
$124,146
Indirect Cost
$9,196
Name
Beth Israel Deaconess Medical Center
Department
Type
DUNS #
071723621
City
Boston
State
MA
Country
United States
Zip Code
02215
Suárez-Fueyo, Abel; Bradley, Sean J; Katsuyama, Takayuki et al. (2018) Downregulation of CD3? in NK Cells from Systemic Lupus Erythematosus Patients Confers a Proinflammatory Phenotype. J Immunol 200:3077-3086
Moulton, Vaishali R; Suarez-Fueyo, Abel; Meidan, Esra et al. (2017) Pathogenesis of Human Systemic Lupus Erythematosus: A Cellular Perspective. Trends Mol Med 23:615-635
Katsuyama, Eri; Yan, Minglu; Watanabe, Katsue Sunahori et al. (2017) Downregulation of miR-200a-3p, Targeting CtBP2 Complex, Is Involved in the Hypoproduction of IL-2 in Systemic Lupus Erythematosus-Derived T Cells. J Immunol 198:4268-4276
Moulton, Vaishali R; Tsokos, George C (2015) T cell signaling abnormalities contribute to aberrant immune cell function and autoimmunity. J Clin Invest 125:2220-7
Moulton, Vaishali R; Gillooly, Andrew R; Perl, Marcel A et al. (2015) Serine Arginine-Rich Splicing Factor 1 (SRSF1) Contributes to the Transcriptional Activation of CD3? in Human T Cells. PLoS One 10:e0131073
Moulton, Vaishali R; Gillooly, Andrew R; Tsokos, George C (2014) Ubiquitination regulates expression of the serine/arginine-rich splicing factor 1 (SRSF1) in normal and systemic lupus erythematosus (SLE) T cells. J Biol Chem 289:4126-34
Moulton, Vaishali R; Grammatikos, Alexandros P; Fitzgerald, Lisa M et al. (2013) Splicing factor SF2/ASF rescues IL-2 production in T cells from systemic lupus erythematosus patients by activating IL-2 transcription. Proc Natl Acad Sci U S A 110:1845-50
Moulton, Vaishali R; Lo, Mindy S; Tsokos, George C (2012) Methods and protocols to study T cell signaling abnormalities in human systemic lupus erythematosus. Methods Mol Biol 900:25-60
Moulton, Vaishali R; Tsokos, George C (2012) Why do women get lupus? Clin Immunol 144:53-6