This renewal continues our optimization of myeloid-derived suppressor cell (MDSC) infusion for creating transplant tolerance to prevent graft-vs-host disease (GVHD) in the clinic. Classical immunosuppressants do not inhibit the initial, innate response critical for tissue inflammation. Myeloid-derived suppressor cells (MDSCs) are innate immune suppressors, functioning by fundamentally different mechanisms, which we hypothesize can suppress the early innate immune cell activation and may participate in tissue injury repair. We showed that highly suppressive murine MDSCs could be generated in short-term culture of normal bone marrow and that monocytic MDSCs (M-MDSCs) were the most potent subset in suppressing GVHD. Because MDSCs can be antigen-independent suppressors, off-the-shelf MDSC preparations for clinical trial(s) are practical. A limitation in translation has been the inherent batch-to-batch variability of primary donor-derived MDSCs. To solve batch variability, we will use transgene-free, clone-based induced pluripotent stem cells (IPSCs) that can be differentiated into suppressive M-MDSCs in 19 days, expandable by 1000-fold, and have phenotypic and functional characteristics of healthy donor peripheral blood MDSCs. This differentiation system was developed by our grant collaborators at Fate Therapeutics who have worked with us/our program to export protocols and a preselected IPSC clone to optimize GMP methods for a planned 2018 IPSC NK cancer trial. Unexpectedly, the greatest remaining impediment is MDSC propensity to terminally differentiate into mature non-suppressive antigen-presenting cells in the inflammatory GVHD milieu, also seen in inflammasome activated human primary MDSCs. With our grant collaborators, we systematically identified key molecular targets to overcome these limitations. Our scientific premise is gene engineering of indefinitely self-renewing IPSCs will subvert inherent MDSC instability in an inflammatory milieu and prolong their in vivo lifespan, creating a single, uniform well-characterized off-the-shelf M-MDSC batch meeting pre- release criteria. We will test the hypotheses that:
Aim 1. Human IPSC or iCD34 gene engineering will provide off-the-shelf, third-party M-MDSCs with optimized antigen-independent GVHD suppression by CRISPR/Cas9 gene disruption of pathways controlling inflammasome activation (NLRP3, shared ASC subunit; ATP receptors P2x7R, P2Y14), inflammatory monocyte differentiation (batf3), and monocyte to macrophage conversion (NFKB1 p50). To prolong in vivo lifespan, we will test c-FLIP overexpression in M-MDSCs. Robust in vitro and in vivo testing in xenogenic GVHD mice will assess key suppression mechanisms and rank potency.
Aim 2. Human anti-AML specific T cell killing of acute myeloid leukemia (AML) blasts will be unimpaired using gene engineered M-MDSCs that preferentially home to inflammation sites. By pinpointing key pathways for M-MDSC viability and suppression and harnessing IPSC technology, our expert team will gain mechanistic insights into controlling GVHD and preserving anti-AML specific T cell responses, providing a platform for GVHD trials.

Public Health Relevance

Our team of experts will develop novel approaches and gain biological insights into immune system control by myeloid-derived suppressor cells (MDSCs). Our findings will have broad implications for the use of MDSCs in controlling adverse immune responses leading to translational applications to harness the full power of MDSCs for hematopoietic stem cell and solid-organ transplantation as well as autoimmunity settings.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL056067-23
Application #
9658658
Study Section
Cancer Immunopathology and Immunotherapy Study Section (CII)
Program Officer
El Kassar, Nahed
Project Start
1995-08-01
Project End
2022-12-31
Budget Start
2019-01-01
Budget End
2019-12-31
Support Year
23
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Pediatrics
Type
Schools of Medicine
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Sarhan, Dhifaf; Hippen, Keli L; Lemire, Amanda et al. (2018) Adaptive NK Cells Resist Regulatory T-cell Suppression Driven by IL37. Cancer Immunol Res 6:766-775
Li, Yue; Guan, Xiaoqun; Liu, Weiren et al. (2018) Helminth-Induced Production of TGF-? and Suppression of Graft-versus-Host Disease Is Dependent on IL-4 Production by Host Cells. J Immunol 201:2910-2922
Li, Yue; Liu, Weiren; Guan, Xiaqun et al. (2018) STAT6 and Furin Are Successive Triggers for the Production of TGF-? by T Cells. J Immunol 201:2612-2623
Bachanova, Veronika; Sarhan, Dhifaf; DeFor, Todd E et al. (2018) Haploidentical natural killer cells induce remissions in non-Hodgkin lymphoma patients with low levels of immune-suppressor cells. Cancer Immunol Immunother 67:483-494
Blazar, Bruce R; MacDonald, Kelli P A; Hill, Geoffrey R (2018) Immune regulatory cell infusion for graft-versus-host disease prevention and therapy. Blood 131:2651-2660
Zitzer, Nina C; Snyder, Katiri; Meng, Xiamoei et al. (2018) MicroRNA-155 Modulates Acute Graft-versus-Host Disease by Impacting T Cell Expansion, Migration, and Effector Function. J Immunol 200:4170-4179
Hülsdünker, Jan; Ottmüller, Katja J; Neeff, Hannes P et al. (2018) Neutrophils provide cellular communication between ileum and mesenteric lymph nodes at graft-versus-host disease onset. Blood 131:1858-1869
Zhang, Ping; Lee, Jason S; Gartlan, Kate H et al. (2017) Eomesodermin promotes the development of type 1 regulatory T (TR1) cells. Sci Immunol 2:
Koehn, Brent H; Blazar, Bruce R (2017) Role of myeloid-derived suppressor cells in allogeneic hematopoietic cell transplantation. J Leukoc Biol 102:335-341
Varelias, Antiopi; Ormerod, Kate L; Bunting, Mark D et al. (2017) Acute graft-versus-host disease is regulated by an IL-17-sensitive microbiome. Blood 129:2172-2185

Showing the most recent 10 out of 167 publications