Background The CAR T cell therapy initiated as a treatment for CD19+ malignant B cell has rapidly expanded to many other cancers including solid tumors. Among multiple key factors to improve efficacy while minimizing adverse effects, it is critical to understand the mechanisms of T cell exhaustion and harness it. Upregulation of TIGIT is a hallmark of exhausted T cells in non-responders among non-Hodgkin’s lymphoma (NHL) patients under CD19+ CAR T therapy.1 However, we have not paid attention to PBMC that can (in)directly interact with CAR+T cells and its synergistic role in the treatment result. Furthermore, it is desirable to identify transcriptional factors (TFs) facilitating the unexpected T cell status. Ultimately, we can target them with TIGIT for better outcomes.
Methods We evaluate 27 scRNA-seq data for CAR+T (n=12) and its matched PBMC (n=15) in two sequential time points after the infusion from 8 relapsed/refractory NHL patients; 6 responders (R) and 2 non-responders (NR).1
Results TIGIT is upregulated in post-infusion PBMC samples. The TIGIT is overexpressed across CD8+T, NK, and most significantly Treg population. DEG analysis reveals that more NR cells activate TIGIT whereas more R cells activate CD226. Furthermore, NECTIN2 is mostly expressed in a monocyte subpopulation dominated by the NR group. CellphoneDB analysis3 demonstrates that KLRC1 in an inflammatory T population in CAR+T interacts with HLA-E detected across PBMC cells (potentially originated from malignant B cells). These suggest that more prevalent immune checkpoints in NR trigger immune evasion.4–7 Next, we investigate how the dysfunctional T state was acquired. A gene set analysis uncovers that it is due to chronic antigen exposure (CAE).2 NK-like T cell populations resemble dysfunctional tumor-infiltrated lymphocytes (TILs) under CAE. Among monocyte populations, the NR-dominant monocytes upregulate TFs represented into CAE TILs. A comprehensive TF analysis reveals that TSC22D3, ZFP36, and DUSP1 known for anti-inflammatory response8 are strikingly overexpressed in the NR group. A higher interaction of TSC22D3 with AP-1 proteins results in a lack of AP-1 to form a binding with NFAT required for T cell effectors. Even worse, NR patients upregulate NR4A2 expression. The gene potentially interferes with the binding of NFAT/AP-1 and remodels chromatin accessibility to facilitate T cell exhaustion.9
Conclusions In conclusion, dysregulated TSC22D3 and NR4A2 compromise the effector function of CAR+T, hijack AP-1 proteins, and impede eradicating the antigens, and thus CAR+T cells transit to exhausted T cells represented by overexpressed TIGIT.
Jackson Z, Hong C, Schauner R, Dropulic B, Caimi PF, de Lima M, Giraudo MF, Gupta K, Reese JS, Hwang TH, Wald DN. Sequential Single-Cell Transcriptional and Protein Marker Profiling Reveals TIGIT as a Marker of CD19 CAR-T Cell Dysfunction in Patients with Non-Hodgkin Lymphoma. Cancer Discov. 2022 Aug 5;12(8):1886–1903. doi: 10.1158/2159–8290.CD-21–1586.
Good CR, Aznar MA, Kuramitsu S, Samareh P, Agarwal S, Donahue G, Ishiyama K, Wellhausen N, Rennels AK, Ma Y, Tian L, Guedan S, Alexander KA, Zhang Z, Rommel PC, Singh N, Glastad KM, Richardson MW, Watanabe K, Tanyi JL, O’Hara MH, Ruella M, Lacey SF, Moon EK, Schuster SJ, Albelda SM, Lanier LL, Young RM, Berger SL, June CH. An NK-like CAR T cell transition in CAR T cell dysfunction. Cell. 2021 Dec 9;184(25):6081–6100.e26. doi: 10.1016/j.cell.2021.11.016. Epub 2021 Dec 2.
Efremova M, Vento-Tormo M, Teichmann SA, Vento-Tormo R. CellPhoneDB: inferring cell-cell communication from combined expression of multi-subunit ligand-receptor complexes. Nat Protoc. 2020 Apr;15(4):1484–1506. doi: 10.1038/s41596–020-0292-x. Epub 2020 Feb 26.
Chiang EY, Mellman I. TIGIT-CD226-PVR axis: advancing immune checkpoint blockade for cancer immunotherapy. J Immunother Cancer. 2022 Apr;10(4):e004711. doi: 10.1136/jitc-2022–004711.
Borst L, van der Burg SH, van Hall T. The NKG2A-HLA-E Axis as a Novel Checkpoint in the Tumor Microenvironment. Clin Cancer Res. 2020 Nov 1;26(21):5549–5556. doi: 10.1158/1078–0432.CCR-19–2095. Epub 2020 May 14.
Odagiu L, May J, Boulet S, Baldwin TA, Labrecque N. Role of the Orphan Nuclear Receptor NR4A Family in T-Cell Biology. Front Endocrinol (Lausanne). 2021 Feb 1;11:624122. doi: 10.3389/fendo.2020.624122.
Mognol GP, Spreafico R, Wong V, Scott-Browne JP, Togher S, Hoffmann A, Hogan PG, Rao A, Trifari S. Exhaustion-associated regulatory regions in CD8+ tumor-infiltrating T cells. Proc Natl Acad Sci U S A. 2017 Mar 28;114(13):E2776-E2785. doi: 10.1073/pnas.1620498114. Epub 2017 Mar 10.
Bereshchenko O, Migliorati G, Bruscoli S, Riccardi C. Glucocorticoid-Induced Leucine Zipper: A Novel Anti-inflammatory Molecule. Front Pharmacol. 2019 Mar 27;10:308. doi: 10.3389/fphar.2019.00308.
Chen J, López-Moyado IF, Seo H, Lio CJ, Hempleman LJ, Sekiya T, Yoshimura A, Scott-Browne JP, Rao A. NR4A transcription factors limit CAR T cell function in solid tumours. Nature. 2019 Mar;567(7749):530–534. doi: 10.1038/s41586–019-0985-x. Epub 2019 Feb 27.
This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.
Statistics from Altmetric.com
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.