Defining inflammatory cell states in rheumatoid arthritis joint synovial tissues by integrating single-cell transcriptomics and mass cytometry
NATURE IMMUNOLOGY
Authors: Zhang, Fan; Wei, Kevin; Slowikowski, Kamil; Fonseka, Chamith Y.; Rao, Deepak A.; Kelly, Stephen; Goodman, Susan M.; Tabechian, Darren; Hughes, Laura B.; Salomon-Escoto, Karen; Watts, Gerald F. M.; Jonsson, A. Helena; Rangel-Moreno, Javier; Meednu, Nida; Rozo, Cristina; Apruzzese, William; Eisenhaure, Thomas M.; Lieb, David J.; Boyle, David L.; Mandelin, Arthur M., II; Boyce, Brendan F.; DiCarlo, Edward; Gravallese, Ellen M.; Gregersen, Peter K.; Moreland, Larry; Firestein, Gary S.; Hacohen, Nir; Nusbaum, Chad; Lederer, James A.; Perlman, Harris; Pitzalis, Costantino; Filer, Andrew; Holers, V. Michael; Bykerk, Vivian P.; Donlin, Laura T.; Anolik, Jennifer H.; Brenner, Michael B.; Raychaudhuri, Soumya; Albrecht, Jennifer; Bridges, S. Louis, Jr.; Buckley, Christopher D.; Buckner, Jane H.; Dolan, James; Guthridge, Joel M.; Gutierrez-Arcelus, Maria; Ivashkiv, Lionel B.; James, Eddie A.; James, Judith A.; Keegan, Josh; Lee, Yvonne C.; McGeachy, Mandy J.; McNamara, Michael A.; Mears, Joseph R.; Mizoguchi, Fumitaka; Nguyen, Jennifer P.; Noma, Akiko; Orange, Dana E.; Rohani-Pichavant, Mina; Ritchlin, Christopher; Robinson, William H.; Seshadri, Anupamaa; Sutherby, Danielle; Seifert, Jennifer; Turner, Jason D.; Utz, Paul J.
Abstract
To define the cell populations that drive joint inflammation in rheumatoid arthritis (RA), we applied single-cell RNA sequencing (scRNA-seq), mass cytometry, bulk RNA sequencing (RNA-seq) and flow cytometry to Tcells, B cells, monocytes, and fibroblasts from 51 samples of synovial tissue from patients with RA or osteoarthritis (OA). Utilizing an integrated strategy based on canonical correlation analysis of 5,265 scRNA-seq profiles, we identified 18 unique cell populations. Combining mass cytometry and transcriptomics revealed cell states expanded in RA synovia: THY1(CD90)(+)HLA-DRA(hi) sublining fibroblasts, IL1B(+) pro-inflammatory monocytes, ITGAX(+)TBX21(+) autoimmune-associated B cells and PDCD1(+) peripheral helper T (T-PH) cells and follicular helper T (T-FH) cells. We defined distinct subsets of CD8(+)Tcells characterized by GZMIK(+), GZMB(+), and GNLY(+) phenotypes. We mapped inflammatory mediators to their source cell populations; for example, we attributed 1L6 expression to THYl(+)HLA-DRA(hi) fibroblasts and IL1B production to pro-inflammatory monocytes. These populations are potentially key mediators of RA pathogenesis.
Myeloid Conditioning with c-kit-Targeted CAR-T Cells Enables Donor Stem Cell Engraftment
MOLECULAR THERAPY
Authors: Arai, Yasuyuki; Choi, Uimook; Corsino, Cristina I.; Koontz, Sherry M.; Tajima, Masaki; Sweeney, Colin L.; Black, Mary A.; Feldman, Steven A.; Dinauer, Mary C.; Malech, Harry L.
Abstract
We report a novel approach to bone marrow (BM) conditioning using c-kit-targeted chimeric antigen receptor T (c-kit CAR-T) cells in mice. Previous reports using anti-c-kit or anti-CD45 antibody linked to a toxin such as saporin have been promising. We developed a distinctly different approach using c-kit CAR-T cells. Initial studies demonstrated in vitro killing of hematopoietic stem cells by c-kit CAR-T cells but poor expansion in vivo and poor migration of CAR-T cells into BM. Pre-treatment of recipient mice with low-dose cyclophosphamide (125 mg/kg) together with CXCR4 transduction in the CAR-T cells enhanced trafficking to and expansion in BM (< 1%-13.1%). This resulted in significant depletion of the BM c-kit(+) population (9.0%-0.1%). Because congenic Thy1.1 CAR-T cells were used in the Thy1.2-recipient mice, anti-Thy1.1 antibody could be used to deplete CAR-T cells in vivo before donor BM transplant. This achieved 20%-40% multilineage engraftment. We applied this conditioning to achieve an average of 28% correction of chronic granulomatous disease mice by wild-type BM transplant. Our findings provide a proof of concept that c-kit CAR-T cells can achieve effective BM conditioning without chemo-/radiotherapy. Our work also demonstrates that co-expression of a trafficking receptor can enhance targeting of CAR-T cells to a designated tissue.
COA
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