The aim of this proposal is to identify and characterize novel long noncoding RNAs (lncRNAs) that function in monocyte to macrophage differentiation. Macrophages are critical effector cells of the innate immune system where they function to control infection and maintain tissue homeostasis. Macrophages are implicated in the pathogenesis of a number of diseases including rheumatoid arthritis and cancer. A critical process for macrophage function involves their differentiation from monocytes present in the peripheral blood. This is a tightly regulated procedure and here we aim to characterize the role that lncRNA play in controlling this process. We will carry out a systematic unbiased screen using Cas9/CRISPRi technology to identify lncRNAs that function to control monocyte to macrophage differentiation. This will allow for rapid meaningful data to be obtained in a highly efficient manner. In our preliminary RNA-sequencing data we have identified hundreds of lncRNA that display differential expression patterns between monocytes and macrophages. We have begun to establish our mechanistic analysis pipeline by investigating the functions of one lncRNA named GAPLINC, which we have found to be induced over 1000 fold during the differentiation process. Here we will employ a suite of in vitro mechanistic techniques to determine how GAPLINC functions to control macrophage differentiation. This project will establish a foundation for future studies to determine the regulatory importance of lncRNAs in innate immune cell development and inflammatory diseases.

Public Health Relevance

In this proposal we will carry out a high throughput unbiased screen to identify long noncoding RNAs (lncRNAs) that function to regulate monocyte to macrophage differentiation. We have data indicating that lncRNAs change expression as macrophages develop and here we will identify and characterize those genes that are critical regulators of this process.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AR070973-01A1
Application #
9387823
Study Section
Cellular and Molecular Immunology - A Study Section (CMIA)
Program Officer
Wang, Yan Z
Project Start
2017-09-01
Project End
2019-08-31
Budget Start
2017-09-01
Budget End
2018-08-31
Support Year
1
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of California Santa Cruz
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
125084723
City
Santa Cruz
State
CA
Country
United States
Zip Code
95064