Complementary CRISPR screen highlights the contrasting role of membrane-bound and soluble ICAM-1 in regulating antigen-specific tumor cell killing by cytotoxic T cells
Cytotoxic CD8 +T lymphocytes (CTLs) are key players of adaptive anti-tumor immunity based on their ability to specifically recognize and destroy tumor cells. Many cancer immunotherapies rely on unleashing CTL function. However, tumors can evade killing through strategies which are not yet fully eluc...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2023-09-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/84314 |
| _version_ | 1797655946937237504 |
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| author | Ann-Kathrin Herzfeldt Marta Puig Gamez Eva Martin Lukasz Miloslaw Boryn Praveen Baskaran Heinrich J Huber Michael Schuler John E Park Lee Kim Swee |
| author_facet | Ann-Kathrin Herzfeldt Marta Puig Gamez Eva Martin Lukasz Miloslaw Boryn Praveen Baskaran Heinrich J Huber Michael Schuler John E Park Lee Kim Swee |
| author_sort | Ann-Kathrin Herzfeldt |
| collection | DOAJ |
| description | Cytotoxic CD8 +T lymphocytes (CTLs) are key players of adaptive anti-tumor immunity based on their ability to specifically recognize and destroy tumor cells. Many cancer immunotherapies rely on unleashing CTL function. However, tumors can evade killing through strategies which are not yet fully elucidated. To provide deeper insight into tumor evasion mechanisms in an antigen-dependent manner, we established a human co-culture system composed of tumor and primary immune cells. Using this system, we systematically investigated intrinsic regulators of tumor resistance by conducting a complementary CRISPR screen approach. By harnessing CRISPR activation (CRISPRa) and CRISPR knockout (KO) technology in parallel, we investigated gene gain-of-function as well as loss-of-function across genes with annotated function in a colon carcinoma cell line. CRISPRa and CRISPR KO screens uncovered 187 and 704 hits, respectively, with 60 gene hits overlapping between both. These data confirmed the role of interferon-γ (IFN-γ), tumor necrosis factor α (TNF-α) and autophagy pathways and uncovered novel genes implicated in tumor resistance to killing. Notably, we discovered that ILKAP encoding the integrin-linked kinase-associated serine/threonine phosphatase 2 C, a gene previously unknown to play a role in antigen specific CTL-mediated killing, mediate tumor resistance independently from regulating antigen presentation, IFN-γ or TNF-α responsiveness. Moreover, our work describes the contrasting role of soluble and membrane-bound ICAM-1 in regulating tumor cell killing. The deficiency of membrane-bound ICAM-1 (mICAM-1) or the overexpression of soluble ICAM-1 (sICAM-1) induced resistance to CTL killing, whereas PD-L1 overexpression had no impact. These results highlight the essential role of ICAM-1 at the immunological synapse between tumor and CTL and the antagonist function of sICAM-1. |
| first_indexed | 2024-03-11T17:22:02Z |
| format | Article |
| id | doaj.art-d6238e319d8e4c0cb86bb5aa4c5e157f |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-03-11T17:22:02Z |
| publishDate | 2023-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-d6238e319d8e4c0cb86bb5aa4c5e157f2023-10-19T10:43:22ZengeLife Sciences Publications LtdeLife2050-084X2023-09-011210.7554/eLife.84314Complementary CRISPR screen highlights the contrasting role of membrane-bound and soluble ICAM-1 in regulating antigen-specific tumor cell killing by cytotoxic T cellsAnn-Kathrin Herzfeldt0https://orcid.org/0000-0002-7882-8284Marta Puig Gamez1https://orcid.org/0009-0006-6047-8200Eva Martin2Lukasz Miloslaw Boryn3https://orcid.org/0000-0001-8091-1071Praveen Baskaran4https://orcid.org/0000-0003-2275-3516Heinrich J Huber5https://orcid.org/0000-0003-4454-2971Michael Schuler6John E Park7https://orcid.org/0000-0002-5674-6026Lee Kim Swee8Department of Cancer Immunology and Immune Modulation, Boehringer Ingelheim, Biberach an der Riss, GermanyDepartment of Cancer Immunology and Immune Modulation, Boehringer Ingelheim, Biberach an der Riss, GermanyDepartment of Drug Discovery Sciences, Boehringer Ingelheim, Biberach an der Riss, GermanyArdigen SA, Kraków, PolandDepartment of Global Computational Biology and Digital Sciences, Boehringer Ingelheim, Biberach an der Riss, GermanyDrug Discovery Sciences, Boehringer Ingelheim, Biberach an der Riss, GermanyDepartment of Drug Discovery Sciences, Boehringer Ingelheim, Biberach an der Riss, GermanyDepartment of Cancer Immunology and Immune Modulation, Boehringer Ingelheim, Biberach an der Riss, GermanyDepartment of Cancer Immunology and Immune Modulation, Boehringer Ingelheim, Biberach an der Riss, GermanyCytotoxic CD8 +T lymphocytes (CTLs) are key players of adaptive anti-tumor immunity based on their ability to specifically recognize and destroy tumor cells. Many cancer immunotherapies rely on unleashing CTL function. However, tumors can evade killing through strategies which are not yet fully elucidated. To provide deeper insight into tumor evasion mechanisms in an antigen-dependent manner, we established a human co-culture system composed of tumor and primary immune cells. Using this system, we systematically investigated intrinsic regulators of tumor resistance by conducting a complementary CRISPR screen approach. By harnessing CRISPR activation (CRISPRa) and CRISPR knockout (KO) technology in parallel, we investigated gene gain-of-function as well as loss-of-function across genes with annotated function in a colon carcinoma cell line. CRISPRa and CRISPR KO screens uncovered 187 and 704 hits, respectively, with 60 gene hits overlapping between both. These data confirmed the role of interferon-γ (IFN-γ), tumor necrosis factor α (TNF-α) and autophagy pathways and uncovered novel genes implicated in tumor resistance to killing. Notably, we discovered that ILKAP encoding the integrin-linked kinase-associated serine/threonine phosphatase 2 C, a gene previously unknown to play a role in antigen specific CTL-mediated killing, mediate tumor resistance independently from regulating antigen presentation, IFN-γ or TNF-α responsiveness. Moreover, our work describes the contrasting role of soluble and membrane-bound ICAM-1 in regulating tumor cell killing. The deficiency of membrane-bound ICAM-1 (mICAM-1) or the overexpression of soluble ICAM-1 (sICAM-1) induced resistance to CTL killing, whereas PD-L1 overexpression had no impact. These results highlight the essential role of ICAM-1 at the immunological synapse between tumor and CTL and the antagonist function of sICAM-1.https://elifesciences.org/articles/84314CRISPRa screenCRISPR KO screentumor cell killingcytotoxic T cellscancer immunologyICAM-1 |
| spellingShingle | Ann-Kathrin Herzfeldt Marta Puig Gamez Eva Martin Lukasz Miloslaw Boryn Praveen Baskaran Heinrich J Huber Michael Schuler John E Park Lee Kim Swee Complementary CRISPR screen highlights the contrasting role of membrane-bound and soluble ICAM-1 in regulating antigen-specific tumor cell killing by cytotoxic T cells eLife CRISPRa screen CRISPR KO screen tumor cell killing cytotoxic T cells cancer immunology ICAM-1 |
| title | Complementary CRISPR screen highlights the contrasting role of membrane-bound and soluble ICAM-1 in regulating antigen-specific tumor cell killing by cytotoxic T cells |
| title_full | Complementary CRISPR screen highlights the contrasting role of membrane-bound and soluble ICAM-1 in regulating antigen-specific tumor cell killing by cytotoxic T cells |
| title_fullStr | Complementary CRISPR screen highlights the contrasting role of membrane-bound and soluble ICAM-1 in regulating antigen-specific tumor cell killing by cytotoxic T cells |
| title_full_unstemmed | Complementary CRISPR screen highlights the contrasting role of membrane-bound and soluble ICAM-1 in regulating antigen-specific tumor cell killing by cytotoxic T cells |
| title_short | Complementary CRISPR screen highlights the contrasting role of membrane-bound and soluble ICAM-1 in regulating antigen-specific tumor cell killing by cytotoxic T cells |
| title_sort | complementary crispr screen highlights the contrasting role of membrane bound and soluble icam 1 in regulating antigen specific tumor cell killing by cytotoxic t cells |
| topic | CRISPRa screen CRISPR KO screen tumor cell killing cytotoxic T cells cancer immunology ICAM-1 |
| url | https://elifesciences.org/articles/84314 |
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