Project Summary/Abstract
The goal of this project is to enhance our understanding of the IDG-eligible kinases DYRK1B, DYRK2,
DYRK3 and DYRK4. Our preliminary data implicate at least one of these kinases in regulating differentiation of
anti-inflammatory Tregs. Treg-modulating therapies are greatly needed, particularly in autoimmunity. We built a
published pipeline to quantitate how genetic and chemical perturbations impact Treg differentiation, this showed
that DYRK1A regulates Th17, but not Treg, differentiation. We now propose to use this pipeline to identify the
Treg-regulating DYRK. This will illuminate a novel immune cellular phenotype for an IDG-eligible gene, develop
validated tools to study IDG-DYRKs in primary cells, inform development of novel Treg-enhancing drugs and
highlight patient subsets for precision therapy. Our long-term goal is to help develop better therapies, including
inhibitors of specific DYRK family members, to treat autoimmunity. The overall objectives in this application are
to (i) develop genetic tools to identify which DYRK family member regulates Treg differentiation, and (ii) interrogate
functional and mechanistic features of DYRK-deficient Tregs. The central hypothesis is that an unidentified DYRK
family member inhibits Treg differentiation. The rationale for this project is that identifying the Treg-regulating DYRK
family member will offer a strong scientific framework to illuminate our understanding of understudied DYRK(s)
as druggable regulators of autoimmunity pathobiology and establish a pipeline to illuminate other IDG-eligible
genes. The central hypothesis will be tested in two specific aims: 1) Define how overexpression of IDG-DYRKs
affects Treg differentiation, and 2) Define how IDG DYRK inhibitors and knockout of IDG-DYRKs affects Treg
differentiation. The first aim will interrogate how overexpressing each DYRK family member in primary murine
and human CD4+ T cells affects Treg differentiation. The second aim will mechanistically interrogate how knockout
of each DYRK family member in primary murine and human CD4+ T cells affects Treg differentiation and function.
These studies leverage our published experimental pipeline that quantitates how genetic or chemical
perturbations impact T cell differentiation. Key innovative features of this proposal include studying the immune
function of IDG-DYRKs and the use of primary immune cells. The proposed research is significant because it is
expected to (i) reveal a new cellular phenotype for an understudied gene of interest to IDG, (ii) identify a novel
druggable regulator of autoimmune pathobiology, thus directing future mechanistic and therapeutic studies, (iii)
develop characterized tools to manipulate understudied DYRKs in other primary cells and biologic contexts and
(iv) establish a scalable pipeline that can be used to rapidly interrogate all genes of interest to IDG for effects on
differentiation into Tregs as well as other lineages including Th17, Th1 and Th2. Ultimately, this has the potential
to broadly illuminate immune function of IDG genes and advance precision therapy of autoimmunity for people
in the general population.