Identifying novel regulators underlying Tregopathies
Rationale and Objectives
Tregs play a particularly critical role in restraining immune responses to self and foreign antigens and their associated inflammation. “Tregopathies” collectively manifest in multiorgan autoimmunity and are caused by loss-of-function mutations in FOXP3, CD25, CTLA4, LRBA, BACH2 and gain-of-function mutations in STAT3. The functional roles of these genes in Tregs have been well studied, which facilitated the identification of mutations in these genes as drivers of “Tregopathies”. However, the spectrum of mechanisms by which Tregs suppress effector T cells and other immune cells is still not fully understood.
1) to perform a genetic screen coupled with an in vitro Treg suppression assay to identify the full spectrum of genes that are involved in Treg-mediated immune suppression.
2) to leverage an arrayed CRISPR/Cas9 knockout library to ablate genes in primary human Tregs.
3) to co-culture in 384 well plates with donor-matched T effector cells in the presence of an activation stimulus. The suppressive effect of Tregs on T effector cell proliferation will be analysed by high throughput flow cytometry. Tregs that lost the ability to suppress effector cells will be analysed and the genes that are ablated in these cells will be considered as hits for in-depth follow up analyses. A comprehensive understanding of genes involved in Treg function will expedite the identification of new inborn errors that cause dysfunction of Tregs. This will be essential to accelerate diagnoses and the design of new targeted therapies to alleviate diseases symptoms or curative approaches that correct the mutations in the genes.
We expect to obtain a comprehensive list of genes that regulate the suppressive function of Tregs. Since mutations in these genes may cause “Tregopathies”, this list of genes will help us to diagnose patients
UKLFR (Grimbacher) to study cells from patients with Tregopathies, m10-13 (3 month); and Alia Therapeutics to learn the standards of genetic manipulation in the biotech industry, m25-27 (2 month).
PhD in Biomedicine, Università delle Svizzera Italiana (Lugano), Switzerland