Anaplastic Lymphoma Kinase (ALK) Inhibitors Enhance Phagocytosis Induced by CD47 Blockade in Sensitive and Resistant ALK-Driven Malignancies
Background: Anaplastic lymphoma kinase (ALK) plays a role in the development of lymphoma, lung cancer and neuroblastoma. While tyrosine kinase inhibitors (TKIs) have improved treatment outcomes, relapse remains a challenge due to on-target mutations and off-target resistance mechanisms. ALK-positive...
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MDPI AG
2024-12-01
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| Online Access: | https://www.mdpi.com/2227-9059/12/12/2819 |
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| author | Federica Malighetti Matteo Villa Mario Mauri Simone Piane Valentina Crippa Ilaria Crespiatico Federica Cocito Elisa Bossi Carolina Steidl Ivan Civettini Chiara Scollo Daniele Ramazzotti Carlo Gambacorti-Passerini Rocco Piazza Luca Mologni Andrea Aroldi |
| author_facet | Federica Malighetti Matteo Villa Mario Mauri Simone Piane Valentina Crippa Ilaria Crespiatico Federica Cocito Elisa Bossi Carolina Steidl Ivan Civettini Chiara Scollo Daniele Ramazzotti Carlo Gambacorti-Passerini Rocco Piazza Luca Mologni Andrea Aroldi |
| author_sort | Federica Malighetti |
| collection | DOAJ |
| description | Background: Anaplastic lymphoma kinase (ALK) plays a role in the development of lymphoma, lung cancer and neuroblastoma. While tyrosine kinase inhibitors (TKIs) have improved treatment outcomes, relapse remains a challenge due to on-target mutations and off-target resistance mechanisms. ALK-positive (ALK+) tumors can evade the immune system, partly through tumor-associated macrophages (TAMs) that facilitate immune escape. Cancer cells use “don’t eat me” signals (DEMs), such as CD47, to resist TAMs-mediated phagocytosis. TKIs may upregulate pro-phagocytic stimuli (i.e., calreticulin, CALR), suggesting a potential therapeutic benefit in combining TKIs with an anti-CD47 monoclonal antibody (mAb). However, the impact of this combination on both TKIs-sensitive and resistant ALK+ tumors requires further investigation. Methods: A panel of TKIs-sensitive and resistant ALK+ cancer subtypes was assessed for CALR and CD47 expression over time using flow cytometry. Flow cytometry co-culture and fluorescent microscopy assays were employed to evaluate phagocytosis under various treatment conditions. Results: ALK inhibitors increased CALR expression in both TKIs-sensitive and off-target resistant ALK+ cancer cells. Prolonged TKIs exposure also led to CD47 upregulation. The combination of ALK inhibitors and anti-CD47 mAb significantly enhanced phagocytosis compared to anti-CD47 alone, as confirmed by flow cytometry and fluorescent microscopy. Conclusions: Anti-CD47 mAb can quench DEMs while exposing pro-phagocytic signals, promoting tumor cell phagocytosis. ALK inhibitors induced immunogenic cell damage by upregulating CALR in both sensitive and off-target resistant tumors. Continuous TKIs exposure in off-target resistant settings also resulted in the upregulation of CD47 over time. Combining TKIs with a CD47 blockade may offer therapeutic benefits in ALK+ cancers, especially in overcoming off-target resistance where TKIs alone are less effective. |
| format | Article |
| id | doaj-art-067420bbdfad46bba2b3a23caffce82d |
| institution | DOAJ |
| issn | 2227-9059 |
| language | English |
| publishDate | 2024-12-01 |
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| series | Biomedicines |
| spelling | doaj-art-067420bbdfad46bba2b3a23caffce82d2025-08-20T02:53:43ZengMDPI AGBiomedicines2227-90592024-12-011212281910.3390/biomedicines12122819Anaplastic Lymphoma Kinase (ALK) Inhibitors Enhance Phagocytosis Induced by CD47 Blockade in Sensitive and Resistant ALK-Driven MalignanciesFederica Malighetti0Matteo Villa1Mario Mauri2Simone Piane3Valentina Crippa4Ilaria Crespiatico5Federica Cocito6Elisa Bossi7Carolina Steidl8Ivan Civettini9Chiara Scollo10Daniele Ramazzotti11Carlo Gambacorti-Passerini12Rocco Piazza13Luca Mologni14Andrea Aroldi15Department of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyDepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyDepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyDepartment of Pathology, Boston Children’s Hospital and Harvard Medical School, Boston, MA 02115, USADepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyDepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyHematology Division, Fondazione IRCCS San Gerardo dei Tintori, 20900 Monza, ItalyHematology Division, Fondazione IRCCS San Gerardo dei Tintori, 20900 Monza, ItalyLymphoma Unit, Department of Onco-Hematology, IRCCS San Raffaele Scientific Institute, 20132 Milan, ItalyExperimental Immunology Unit, IRCCS San Raffaele Scientific Institute, 20132 Milan, ItalyTransfusion Medicine Unit, Fondazione IRCCS San Gerardo dei Tintori, 20900 Monza, ItalyDepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyDepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyDepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyDepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyDepartment of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, ItalyBackground: Anaplastic lymphoma kinase (ALK) plays a role in the development of lymphoma, lung cancer and neuroblastoma. While tyrosine kinase inhibitors (TKIs) have improved treatment outcomes, relapse remains a challenge due to on-target mutations and off-target resistance mechanisms. ALK-positive (ALK+) tumors can evade the immune system, partly through tumor-associated macrophages (TAMs) that facilitate immune escape. Cancer cells use “don’t eat me” signals (DEMs), such as CD47, to resist TAMs-mediated phagocytosis. TKIs may upregulate pro-phagocytic stimuli (i.e., calreticulin, CALR), suggesting a potential therapeutic benefit in combining TKIs with an anti-CD47 monoclonal antibody (mAb). However, the impact of this combination on both TKIs-sensitive and resistant ALK+ tumors requires further investigation. Methods: A panel of TKIs-sensitive and resistant ALK+ cancer subtypes was assessed for CALR and CD47 expression over time using flow cytometry. Flow cytometry co-culture and fluorescent microscopy assays were employed to evaluate phagocytosis under various treatment conditions. Results: ALK inhibitors increased CALR expression in both TKIs-sensitive and off-target resistant ALK+ cancer cells. Prolonged TKIs exposure also led to CD47 upregulation. The combination of ALK inhibitors and anti-CD47 mAb significantly enhanced phagocytosis compared to anti-CD47 alone, as confirmed by flow cytometry and fluorescent microscopy. Conclusions: Anti-CD47 mAb can quench DEMs while exposing pro-phagocytic signals, promoting tumor cell phagocytosis. ALK inhibitors induced immunogenic cell damage by upregulating CALR in both sensitive and off-target resistant tumors. Continuous TKIs exposure in off-target resistant settings also resulted in the upregulation of CD47 over time. Combining TKIs with a CD47 blockade may offer therapeutic benefits in ALK+ cancers, especially in overcoming off-target resistance where TKIs alone are less effective.https://www.mdpi.com/2227-9059/12/12/2819ALKmacrophagestumor immunologyCD47TKIs (tyrosine kinase inhibitors)neuroblastoma |
| spellingShingle | Federica Malighetti Matteo Villa Mario Mauri Simone Piane Valentina Crippa Ilaria Crespiatico Federica Cocito Elisa Bossi Carolina Steidl Ivan Civettini Chiara Scollo Daniele Ramazzotti Carlo Gambacorti-Passerini Rocco Piazza Luca Mologni Andrea Aroldi Anaplastic Lymphoma Kinase (ALK) Inhibitors Enhance Phagocytosis Induced by CD47 Blockade in Sensitive and Resistant ALK-Driven Malignancies Biomedicines ALK macrophages tumor immunology CD47 TKIs (tyrosine kinase inhibitors) neuroblastoma |
| title | Anaplastic Lymphoma Kinase (ALK) Inhibitors Enhance Phagocytosis Induced by CD47 Blockade in Sensitive and Resistant ALK-Driven Malignancies |
| title_full | Anaplastic Lymphoma Kinase (ALK) Inhibitors Enhance Phagocytosis Induced by CD47 Blockade in Sensitive and Resistant ALK-Driven Malignancies |
| title_fullStr | Anaplastic Lymphoma Kinase (ALK) Inhibitors Enhance Phagocytosis Induced by CD47 Blockade in Sensitive and Resistant ALK-Driven Malignancies |
| title_full_unstemmed | Anaplastic Lymphoma Kinase (ALK) Inhibitors Enhance Phagocytosis Induced by CD47 Blockade in Sensitive and Resistant ALK-Driven Malignancies |
| title_short | Anaplastic Lymphoma Kinase (ALK) Inhibitors Enhance Phagocytosis Induced by CD47 Blockade in Sensitive and Resistant ALK-Driven Malignancies |
| title_sort | anaplastic lymphoma kinase alk inhibitors enhance phagocytosis induced by cd47 blockade in sensitive and resistant alk driven malignancies |
| topic | ALK macrophages tumor immunology CD47 TKIs (tyrosine kinase inhibitors) neuroblastoma |
| url | https://www.mdpi.com/2227-9059/12/12/2819 |
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