Identification of TUBB3 as an immunotherapy target in lung cancer by genome wide in vivo CRISPR screening
Recent development of immune checkpoint inhibitors has revolutionized cancer immunotherapy. Although these drugs show dramatic effects on a subset of cancer patients, many other tumors are non-responsive and the pathological mechanism of the resistance is largely unknown. To identify genes underlyin...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-02-01
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| Series: | Neoplasia: An International Journal for Oncology Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S1476558624001416 |
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| author | Dan Zhao Ravindra Deshpande Kerui Wu Abhishek Tyagi Sambad Sharma Shih-Ying Wu Fei Xing Stacey O'Neill Jimmy Ruiz Feng Lyu Kounosuke Watabe |
| author_facet | Dan Zhao Ravindra Deshpande Kerui Wu Abhishek Tyagi Sambad Sharma Shih-Ying Wu Fei Xing Stacey O'Neill Jimmy Ruiz Feng Lyu Kounosuke Watabe |
| author_sort | Dan Zhao |
| collection | DOAJ |
| description | Recent development of immune checkpoint inhibitors has revolutionized cancer immunotherapy. Although these drugs show dramatic effects on a subset of cancer patients, many other tumors are non-responsive and the pathological mechanism of the resistance is largely unknown. To identify genes underlying anti-PD-1 immunotherapy resistance using a systematic approach, we performed an in vivo genome wide CRISPR screening in lung cancer cells. We integrated our results with multi-omics clinical data and performed both in vitro and in vivo assays to evaluate the role of the top candidate in regulating cytotoxic T cell killing. We identified TUBB3 as a potential target to overcome the resistance and enhance the efficacy of anti-PD-1 immunotherapy. TUBB3 expression is upregulated in lung cancer patients, and its higher expression correlates with poorer patients’ survival. We found that TUBB3 expression was significantly elevated in the non-responders compared to responders in our patient cohort that received immunotherapies. Importantly, the results of our preclinical experiments showed that inhibition of TUBB3 with a small molecule inhibitor synergized with anti-PD-1 treatment and enhanced tumor cell killing by cytotoxic T cells. Consistently, anti-PD-1 resistant cells showed significantly higher expression of TUBB3; however, TUBB3 inhibition rendered the resistant cells more susceptible to T cell killing. Mechanistic studies revealed that blocking TUBB3 suppressed the expression of PD-L1 through the EMT-related SNAI1 gene. Our results provide a rationale for a novel combination therapy consisting of the TUBB3 inhibition and anti-PD-1 immunotherapy for lung cancer. |
| format | Article |
| id | doaj-art-091b6cd2a848416da425f9bc2ee72f73 |
| institution | Kabale University |
| issn | 1476-5586 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neoplasia: An International Journal for Oncology Research |
| spelling | doaj-art-091b6cd2a848416da425f9bc2ee72f732025-02-03T04:16:32ZengElsevierNeoplasia: An International Journal for Oncology Research1476-55862025-02-0160101100Identification of TUBB3 as an immunotherapy target in lung cancer by genome wide in vivo CRISPR screeningDan Zhao0Ravindra Deshpande1Kerui Wu2Abhishek Tyagi3Sambad Sharma4Shih-Ying Wu5Fei Xing6Stacey O'Neill7Jimmy Ruiz8Feng Lyu9Kounosuke Watabe10Department of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USA; Baylor College of Medicine, 1 Baylor Plz, Houston, TX 77030, USADepartment of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USAUniversity of North Carolina, Greensboro, NC, 27412, USADepartment of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USAAuron Therapeutics, Massachusetts, 02458, USADepartment of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USADepartment of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USACWRU School of Medicine, Cleveland, OH 44106-7288, USADepartment of Medicine (Hematology & Oncology), Wake Forest School of Medicine, Winston Salem, NC 27157, USADepartment of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USADepartment of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USA; Corresponding author at: Department of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USA.Recent development of immune checkpoint inhibitors has revolutionized cancer immunotherapy. Although these drugs show dramatic effects on a subset of cancer patients, many other tumors are non-responsive and the pathological mechanism of the resistance is largely unknown. To identify genes underlying anti-PD-1 immunotherapy resistance using a systematic approach, we performed an in vivo genome wide CRISPR screening in lung cancer cells. We integrated our results with multi-omics clinical data and performed both in vitro and in vivo assays to evaluate the role of the top candidate in regulating cytotoxic T cell killing. We identified TUBB3 as a potential target to overcome the resistance and enhance the efficacy of anti-PD-1 immunotherapy. TUBB3 expression is upregulated in lung cancer patients, and its higher expression correlates with poorer patients’ survival. We found that TUBB3 expression was significantly elevated in the non-responders compared to responders in our patient cohort that received immunotherapies. Importantly, the results of our preclinical experiments showed that inhibition of TUBB3 with a small molecule inhibitor synergized with anti-PD-1 treatment and enhanced tumor cell killing by cytotoxic T cells. Consistently, anti-PD-1 resistant cells showed significantly higher expression of TUBB3; however, TUBB3 inhibition rendered the resistant cells more susceptible to T cell killing. Mechanistic studies revealed that blocking TUBB3 suppressed the expression of PD-L1 through the EMT-related SNAI1 gene. Our results provide a rationale for a novel combination therapy consisting of the TUBB3 inhibition and anti-PD-1 immunotherapy for lung cancer.http://www.sciencedirect.com/science/article/pii/S1476558624001416In vivo CRISPR screeningTUBB3Anti-PD-1PDL-1Lung cancer immunotherapy |
| spellingShingle | Dan Zhao Ravindra Deshpande Kerui Wu Abhishek Tyagi Sambad Sharma Shih-Ying Wu Fei Xing Stacey O'Neill Jimmy Ruiz Feng Lyu Kounosuke Watabe Identification of TUBB3 as an immunotherapy target in lung cancer by genome wide in vivo CRISPR screening Neoplasia: An International Journal for Oncology Research In vivo CRISPR screening TUBB3 Anti-PD-1 PDL-1 Lung cancer immunotherapy |
| title | Identification of TUBB3 as an immunotherapy target in lung cancer by genome wide in vivo CRISPR screening |
| title_full | Identification of TUBB3 as an immunotherapy target in lung cancer by genome wide in vivo CRISPR screening |
| title_fullStr | Identification of TUBB3 as an immunotherapy target in lung cancer by genome wide in vivo CRISPR screening |
| title_full_unstemmed | Identification of TUBB3 as an immunotherapy target in lung cancer by genome wide in vivo CRISPR screening |
| title_short | Identification of TUBB3 as an immunotherapy target in lung cancer by genome wide in vivo CRISPR screening |
| title_sort | identification of tubb3 as an immunotherapy target in lung cancer by genome wide in vivo crispr screening |
| topic | In vivo CRISPR screening TUBB3 Anti-PD-1 PDL-1 Lung cancer immunotherapy |
| url | http://www.sciencedirect.com/science/article/pii/S1476558624001416 |
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