Related Experiment Video
Updated: Jul 28, 2025
In Vivo CRISPR/Cas9 Screening to Simultaneously Evaluate Gene Function in Mouse Skin and Oral Cavity
Published on: November 2, 2020
In vivo CRISPR screens reveal Serpinb9 and Adam2 as regulators of immune therapy response in lung cancer
Dzana Dervovic1, Ahmad A Malik1,2, Edward L Y Chen1
1Centre for Molecular and Systems Biology, Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada.
Abstract:
How the genetic landscape governs a tumor's response to immunotherapy remains poorly understood. To assess the immune-modulatory capabilities of 573 genes associated with altered cytotoxicity in human cancers, here we perform CRISPR/Cas9 screens directly in mouse lung cancer models. We recover the known immune evasion factors Stat1 and Serpinb9 and identify the cancer testis antigen Adam2 as an immune modulator, whose expression is induced by KrasG12D and further elevated by immunotherapy. Using loss- and gain-of-function experiments, we show that ADAM2 functions as an oncogene by restraining interferon and TNF cytokine signaling causing reduced presentation of tumor-associated antigens. ADAM2 also restricts expression of the immune checkpoint inhibitors PDL1, LAG3, TIGIT and TIM3 in the tumor microenvironment, which might explain why ex vivo expanded and adoptively transferred cytotoxic T-cells show enhanced cytotoxic efficacy in ADAM2 overexpressing tumors. Together, direct in vivo CRISPR/Cas9 screens can uncover genetic alterations that control responses to immunotherapies.
Insights
The study identified ADAM2 as a key gene influencing tumor response to immunotherapy. Its expression hinders anti-tumor immunity by suppressing cytokine signaling and immune checkpoints.
Area of Science:
- Oncology
- Immunology
- Genetics
Background:
- Understanding the genetic basis of tumor response to immunotherapy is crucial for developing effective cancer treatments.
- Identifying genes that modulate anti-tumor immunity can reveal new therapeutic targets.
Purpose of the Study:
- To investigate the role of 573 genes in tumor immune modulation using in vivo CRISPR screens.
- To identify novel genetic factors governing a tumor's response to immunotherapy.
Main Methods:
- Conducted CRISPR/Cas9 screens in mouse lung cancer models to assess gene function in immune modulation.
- Performed loss- and gain-of-function experiments to validate identified genes, including ADAM2.
Main Results:
- Identified ADAM2, a cancer-testis antigen, as a novel immune modulator.
- Demonstrated that ADAM2 restrains interferon and TNF signaling, reducing tumor-associated antigen presentation.
- Showed ADAM2 restricts immune checkpoint inhibitor expression (PDL1, LAG3, TIGIT, TIM3), enhancing T-cell efficacy.
Conclusions:
- In vivo CRISPR screens are effective for discovering genetic regulators of immunotherapy response.
- ADAM2 acts as an oncogene that suppresses anti-tumor immunity and can be a potential therapeutic target.
