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CAR T cells are remarkably effective in hematologic malignancies, but tumor cells expressing low antigen levels can escape elimination. In Nature Medicine, Mansilla-Soto et al. design new chimeric receptors that link the variable regions of antibodies directly to T cell receptor chains and recognize tumor cells with improved antigen sensitivity., Competing Interests: Declaration of interests S.R.R. is a founder of Juno Therapeutics, now Juno/Bristol Myers Squibb, and a founder of and advisor to Lyell Immunopharma. S.R.R. has patents related to the field of chimeric antigen receptor-modified T cells and their use in immunotherapy., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The TMC genes encode a set of homologous transmembrane proteins whose functions are not well understood. Biallelic mutations in either TMC6 or TMC8 are detected in more than half of cases of the pre-malignant skin disease epidermodysplasia verruciformis (EV). It is controversial whether EV induced by mutations in TMC6 or TMC8 originates from keratinocyte or lymphocyte defects. Quantification of TMC6 and TMC8 RNA levels in various organs revealed that lymphoid tissues have the highest levels of expression of both genes, and custom antibodies confirmed protein expression in mouse lymphocytes. To study the function of these proteins we generated mice with targeted deletion mutant alleles of Tmc6 or Tmc8 Either TMC6 or TMC8 deficiency induced a reduction in apparent molecular weight and/or amount of the other TMC molecule. Co-immunoprecipitation experiments indicated that TMC6 and TMC8 formed a protein complex in mouse and human T cells. MS and biochemical analysis demonstrated that TMC6 and TMC8 additionally interacted with the CIB1 protein to form TMC6-TMC8-CIB1 trimers. We demonstrated that TMC6 and TMC8 regulated CIB1 levels by protecting CIB1 from ubiquitination and proteasomal degradation. Reciprocally, CIB1 was needed for stabilizing TMC6 and TMC8 levels. These results suggest why inactivating mutations in any of the three human genes leads to similar clinical presentations. We also demonstrated that TMC6 and TMC8 levels are drastically lower and the proteins are less active in regulating CIB1 in keratinocytes than in T cells. Our study suggests that defects in lymphocytes may contribute to the etiology and pathogenesis of EV., Competing Interests: Conflict of interest—The authors declare that they have no conflicts of interest with the contents of this article.

By including oligonucleotide-labeled antibodies into high-throughput single-cell RNA-sequencing protocols, combined transcript and protein expression data can be acquired on the single-cell level. Here, we describe a protocol for the combined analysis of over 40 proteins and 400 genes on over 10 4 cells using the nano-well based Rhapsody platform. We also include a workflow for sample multiplexing, which uniquely identifies the initial source of cells (such as tissue type or donor) in the downstream analysis after upstream pooling. For complete information on the use and execution of this protocol, please refer to Mair et al. (2020)., Competing Interests: DECLARATION OF INTERESTS J.R.E., F.M. G.B., and M.P. declare no competing interests. J.M., A.J.T., S.M., and M.N. are employees of BD Biosciences.