Genetic and Cellular Engineering in Immunology and Regenerative Medicine
Team 2

We aim to integrate immune tolerance, transgenesis, gene editing and human ES/iPS cells to explore and manipulate immune responses

Description

Objectives:

1) Work on CD8+ Tregs, in rat, mice and human, both on basic (phenotype, function, TCR-peptide interactions, CAR, RNA-seq) and clinical aspects (in pathological situations and as cellular therapy product).

2) New therapeutic startegies to induce tolerance: we generated a Mab allowing to deplete CD4+ and CD8+ effector T cells without affecting CD4+ and CD8+ Tregs and a cytokine inducing CD4+ and CD8+ Tregs. We are exploring their role in transplantation and autoimmune diseases.

3) The generation of KO and KI rats for immune genes (such as Ighm, Rag1 and Il2Rg KO) using nucleases (ZFNs, TALENs, CRISPRs) through an IBiSA-labeled facility. We generate CRISPRs to analyze the function of new genes, label or ablate specific cell lineages (such as Foxp3) and generate conditional KOs.

4) The analyses of stemness of ES/iPS using single cells from human and rat blastocysts and RNAseq. As a result, new truly naïve human and rat ES cells can be defined and better control their differentiation to other cell types and their application in rat transgenesis.

5) The generation of human GMP hepatocytes derived from ES/iPS cells to transplant in liver diseases. We will also correct by gene-editing iPS cells from patients with liver genetic diseases using CRISPRs and derive hepatocytes from them for personalized regenerative medicine.

6)Differentiation of human ES/iPS towards CD34+ cells and then to CD8+ Tregs. ES/iPS derived CD34+ cells and hepatocytes will be used to humanize immunodeficient rats as new pathophysiological models. ES/iPS-derived CD8+ Tregs will be a source of therapeutic cells.

7)Gene editing of immune genes using CRISPRs and different viral vectors. Immunogene editing will be applied to human CD8+ Tregs to KO or KI genes and to introduce TCRs or CARs directed against alloantigens to give higher specificity to CD8 Tregs. Human CD34+ cells derived from ES/iPS will be KO or KI for different immune genes and then used to humanize immunodeficient mice or rats to explore the function of these genes.