|
| Status |
Public on Oct 16, 2020 |
| Title |
MLLT6 maintains PD‐L1 expression and mediates tumor immune resistance |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Other
|
| Summary |
We present a CRISPR-Cas9 screen in human cancer cells to identify genes that confers tumors with the ability to evade the cytotoxic effects of the immune system. We discovered that the transcriptional regulator MLLT6 (AF17) is required for PD-L1 cell surface presentation and protein expression in multiple cancer types and MLLT6 depletion alleviates suppression of CD8+ cytotoxic T cell-mediated cytolysis. Furthermore, cancer cells lacking MLLT6 exhibit impaired STAT1 signaling and are insensitive to interferon gamma induced stimulation of IDO1, GBP5, CD74 and MHC class II genes, while maintaining MHC class I gene expression. Collectively, our findings establish MLLT6 as a regulator of oncogenic and interferon gamma associated immune resistance
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| |
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| Overall design |
Pooled CRISPR-Cas9 based screen to identify genes regulating PD-L1. Cells that expressed low levels of PD-L1 were subjected to deep sequencing to determine frequencies of sgRNAs and their target genes that were enriched, IFNG treated and untreated samples from 3 different time points were used for studying the sgRNA expression profiles by deepsequencing, RNA-seq transcriptome profiling of U2OS wildtype and MLLT6 knockout cells treated with and without IFNG. Samples of each condition was submitted in triplicates for deep sequencing.
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| Web link |
https://doi.org/10.15252/embr.202050155
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| |
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| Contributor(s) |
Theis M, Sreevalsan S, Paszkowski-Rogacz M |
| Citation(s) |
33063451 |
| |
| Submission date |
Jan 29, 2020 |
| Last update date |
Nov 03, 2020 |
| Contact name |
Maciej Paszkowski-Rogacz |
| Organization name |
Technische Universität Dresden
|
| Department |
UCC / Medical Faculty and University Hospital Carl Gustav Carus
|
| Lab |
Medical Systems Biology (Prof. Buchholz)
|
| Street address |
Tatzberg 47/49
|
| City |
Dresden |
| ZIP/Postal code |
01307 |
| Country |
Germany |
| |
|
| Platforms (1) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
|
| Samples (16)
|
| GSM4289029 |
RKO PD-L1-EGFP reporter line with sgRNA library |
| GSM4289030 |
RKO PD-L1-EGFP reporter line with sgRNA library (viability check) before sort |
| GSM4289031 |
RKO PD-L1-EGFP reporter line with sgRNA library without IFNG treatment |
| GSM4289032 |
RKO PD-L1-EGFP reporter line with sgRNA library with IFNG treatment |
| GSM4289033 |
Mock cells without IFNG - replicate 1 |
| GSM4289034 |
Mock cells without IFNG - replicate 2 |
| GSM4289035 |
Mock cells without IFNG - replicate 3 |
| GSM4289036 |
Mock cells with IFNG - replicate 1 |
| GSM4289037 |
Mock cells with IFNG - replicate 2 |
| GSM4289038 |
Mock cells with IFNG - replicate 3 |
| GSM4289039 |
MLLT6 knockout without IFNG - replicate 1 |
| GSM4289040 |
MLLT6 knockout without IFNG - replicate 2 |
| GSM4289041 |
MLLT6 knockout without IFNG - replicate 3 |
| GSM4289042 |
MLLT6 knockout with IFNG - replicate 1 |
| GSM4289043 |
MLLT6 knockout with IFNG - replicate 2 |
| GSM4289044 |
MLLT6 knockout with IFNG - replicate 3 |
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| Relations |
| BioProject |
PRJNA603767 |
| SRA |
SRP245883 |
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