Zinc finger protein Zfp335 controls early T-cell development and survival through β-selection-dependent and -independent mechanisms

Abstract

T cell development in the thymus undergoes the process of differentiation, selective proliferation and survival from CD4-CD8- double negative (DN) stage to CD4+CD8+ double positive (DP) stage prior to the formation of CD4+ helper and CD8+ cytolytic T cells ready for circulation. Each developmental stage is tightly regulated by sequentially-operating molecular networks, of which only limited numbers of transcription regulators have been deciphered. Here we identified Zfp335 transcription factor as a new player in the regulatory network controlling thymocyte development in mice. We demonstrate that Zfp335 intrinsically controls DN to DP transition, as T cell-specific deficiency in Zfp335 leads to a substantial accumulation of DN3 along with reduction of DP, CD4+ and CD8+ thymocytes. This developmental blockade at DN stage results from the impaired intracellular TCRβ expression as well as increased susceptibility to apoptosis in thymocytes. Transcriptomic and ChIP-seq analyses revealed a direct regulation of transcription factors Bcl6 and Rorcβ by Zfp335. Importantly, enhanced expression of TCRβ and Bcl6/Rorc restores the developmental defect during DN3 to DN4 transition and improves thymocytes survival, respectively. These findings identify a critical role of Zfp335 in controlling T cell development by maintaining intracellular TCRβ expression-mediated β-selection and independently activating cell survival signaling.

Thymically imprinted differential responsivity of naïve CD4 T cells to IL2 results in a heterogeneity of both Treg induction and subsequent effector functioning

“…oxidative phosphorylation [Fig 6f]). We also observed enrichment in genes associated with the transcription factor zinc finger protein 335 (ZFP355, Fig 6g) recently ascribed to have a role in thymic development and regulation of the genes BCL6 and RORC (Wang et al, 2022). Finally, we observed enrichment of genes associated with IL-2 and STAT5 signaling (Fig 6h).…”

Zfp335 establishes eTreg lineage and neonatal immune tolerance by targeting Hadha-mediated fatty acid oxidation

“…Germline knockout of Zfp335 is embryonically lethal, whereas the Zfp335bloto allele, a missense mutation derived from N -ethyl- N -nitrosourea (ENU) mutagenesis, leads to a significant reduction in the number of peripheral T cells due to defects in the maturation and migration of thymocytes ( 46 ). Our previous studies also demonstrate a severe block at the double-negative (DN) stage of thymocyte development upon LckCre-induced Zfp335 deletion ( 47 ). Zfp335 is also involved in memory CD8 + T cell differentiation by its direct regulation of transcription factor 7 (TCF7) expression ( 47 ).…”

Down-regulation of zinc finger protein 335 undermines natural killer cell function in mouse colitis-associated colorectal carcinoma

“…ZFP335 regulates gene transcription by forming complexes with NRC and histone methyltransferase factors or by directly binding DNA [ [16] , [17] , [18] ]. The target genes of ZFP335, such as Bcl6 , Rorc , and Tcf7 , are essential for cell proliferation, cell death, metabolism, mitochondrial function, RNA processing, and transcriptional regulation [ [ 17 , [19] , [20] , [21] ]]. Therefore, ZFP335 down-regulation might elicit multiple biological alterations of NK cells, necessitating future revelation of ZFP335-mediated changes in transcriptomic and proteomic profiles.…”

“…ZFP335 can directly bind to DNA and promote transcription via recognizing two consensus DNA motifs [ 18 ]. Recent research implies that ZFP335 is necessary for transcriptional programming of T cell precursors and the formation of memory CD8 + T cells [ [ [19] , [20] , [21] ]]. However, the importance of ZFP335 for NK cell function remains unidentified.…”