ZFP148 is a transcriptional regulator of effector CD8+ T cell differentiation

In the setting of chronic antigen stimulation, such as chronic viral infection and cancer, antigen-specific CD8+ T cells differentiate into a heterogeneous pool of progenitor/stem-like, effector, and exhausted/dysfunctional subsets. Though genetic and epigenetic programs that contribute to the generation of each subset have been reported, how these subsets are dynamically regulated and how their balance can be shifted from exhausted toward cytolytic effector cells remain as outstanding questions. Here, we identified Zinc-Finger Protein 148 (ZFP148) as a novel transcriptional checkpoint for CD8+ T cell effector differentiation. We performed CUT&Tag followed by seuqencing (CUT&Tag-seq) of ZFP148 in activated CD8+ T cells from healthy C57BL/6J mice to identify direct DNA-binding activity of ZFP148. Overall design: Naïve CD8+ T cells were isolated from the spleens of healthy mice and activated with 5 µg/ml plate-bound anti-CD3 and 2 µg/ml soluble anti-CD28 antibody for 24 hours in 24-well plate. A total of 1 million cells were used for library construction of each target (ZFP148 or IgG control) using the CUT&Tag-seq following the stepwise protocol from Epicypher https://www.epicypher.com/resources/protocols/cutana-cut-and-tag-protocol/. Libraries with equal molar ratio were pooled together. Paired-end sequencing was performed on an Illumina Novaseq X plus platform by Azenta Life Sciences, and 50 million reads were generated for each sample.