Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockade
Background Tigilanol tiglate (TT) is a protein kinase C (PKC)/C1 domain activator currently being developed as an intralesional agent for the treatment of various (sub)cutaneous malignancies. Previous work has shown that intratumoral (I.T.) injection of TT causes vascular disruption with concomitant...
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BMJ Publishing Group
2024-04-01
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| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/12/4/e006602.full |
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| author | Kevin Hendrawan Kelly M Brooks Pei-Yi Yap Blake Ferguson Motoko Koyama Herlina Handoko Jenny Johns Natasa Broit Praphaporn Stewart Daniel Shelley Tracey McMahon Yi Chieh Lim Giovanni Appendino Jason K Cullen Zara C Bruce Steven M Ogbourne Paul W Reddell Glen M Boyle Peter G Parsons Jacinta L Simmons Emily S Wilson Marjorie M A de Souza Tam Hong Nguyen Alberto Pagani Victoria A Gordon |
| author_facet | Kevin Hendrawan Kelly M Brooks Pei-Yi Yap Blake Ferguson Motoko Koyama Herlina Handoko Jenny Johns Natasa Broit Praphaporn Stewart Daniel Shelley Tracey McMahon Yi Chieh Lim Giovanni Appendino Jason K Cullen Zara C Bruce Steven M Ogbourne Paul W Reddell Glen M Boyle Peter G Parsons Jacinta L Simmons Emily S Wilson Marjorie M A de Souza Tam Hong Nguyen Alberto Pagani Victoria A Gordon |
| author_sort | Kevin Hendrawan |
| collection | DOAJ |
| description | Background Tigilanol tiglate (TT) is a protein kinase C (PKC)/C1 domain activator currently being developed as an intralesional agent for the treatment of various (sub)cutaneous malignancies. Previous work has shown that intratumoral (I.T.) injection of TT causes vascular disruption with concomitant tumor ablation in several preclinical models of cancer, in addition to various (sub)cutaneous tumors presenting in the veterinary clinic. TT has completed Phase I dose escalation trials, with some patients showing signs of abscopal effects. However, the exact molecular details underpinning its mechanism of action (MoA), together with its immunotherapeutic potential in oncology remain unclear.Methods A combination of microscopy, luciferase assays, immunofluorescence, immunoblotting, subcellular fractionation, intracellular ATP assays, phagocytosis assays and mixed lymphocyte reactions were used to probe the MoA of TT in vitro. In vivo studies with TT used MM649 xenograft, CT-26 and immune checkpoint inhibitor refractory B16-F10-OVA tumor bearing mice, the latter with or without anti-programmed cell death 1 (PD-1)/anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) mAb treatment. The effect of TT at injected and non-injected tumors was also assessed.Results Here, we show that TT induces the death of endothelial and cancer cells at therapeutically relevant concentrations via a caspase/gasdermin E-dependent pyroptopic pathway. At therapeutic doses, our data demonstrate that TT acts as a lipotoxin, binding to and promoting mitochondrial/endoplasmic reticulum (ER) dysfunction (leading to unfolded protein responsemt/ER upregulation) with subsequent ATP depletion, organelle swelling, caspase activation, gasdermin E cleavage and induction of terminal necrosis. Consistent with binding to ER membranes, we found that TT treatment promoted activation of the integrated stress response together with the release/externalization of damage-associated molecular patterns (HMGB1, ATP, calreticulin) from cancer cells in vitro and in vivo, characteristics indicative of immunogenic cell death (ICD). Confirmation of ICD in vivo was obtained through vaccination and rechallenge experiments using CT-26 colon carcinoma tumor bearing mice. Furthermore, TT also reduced tumor volume, induced immune cell infiltration, as well as improved survival in B16-F10-OVA tumor bearing mice when combined with immune checkpoint blockade.Conclusions These data demonstrate that TT is an oncolytic small molecule with multiple targets and confirms that cell death induced by this compound has the potential to augment antitumor responses to immunotherapy. |
| format | Article |
| id | doaj-art-756c2bca74d94c10bc6e590de097d302 |
| institution | Kabale University |
| issn | 2051-1426 |
| language | English |
| publishDate | 2024-04-01 |
| publisher | BMJ Publishing Group |
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| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj-art-756c2bca74d94c10bc6e590de097d3022024-11-14T01:40:07ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262024-04-0112410.1136/jitc-2022-006602Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockadeKevin Hendrawan0Kelly M Brooks1Pei-Yi Yap2Blake Ferguson3Motoko Koyama4Herlina Handoko5Jenny Johns6Natasa Broit7Praphaporn Stewart8Daniel Shelley9Tracey McMahon10Yi Chieh Lim11Giovanni Appendino12Jason K Cullen13Zara C Bruce14Steven M Ogbourne15Paul W Reddell16Glen M Boyle17Peter G Parsons18Jacinta L Simmons19Emily S Wilson20Marjorie M A de Souza21Tam Hong Nguyen22Alberto Pagani23Victoria A Gordon24QIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaUniversity of the Sunshine Coast, Maroochydore DC, Queensland, AustraliaUniversity of the Sunshine Coast, Maroochydore DC, Queensland, AustraliaUniversity of the Sunshine Coast, Maroochydore DC, Queensland, AustraliaDanish Cancer Society Research Centre, Copenhagen DK, DenmarkDipartimento di Scienze del Farmaco, Università Degli Studi del Piemonte Orientale, Novara, ItalyQBiotics Group Limited, Brisbane, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQBiotics Group Limited, Brisbane, Queensland, AustraliaQBiotics Group Limited, Brisbane, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaQIMR Berghofer Medical Research Institute, Herston, Queensland, AustraliaDipartimento di Scienze del Farmaco, Università Degli Studi del Piemonte Orientale, Novara, ItalyQBiotics Group Limited, Brisbane, Queensland, AustraliaBackground Tigilanol tiglate (TT) is a protein kinase C (PKC)/C1 domain activator currently being developed as an intralesional agent for the treatment of various (sub)cutaneous malignancies. Previous work has shown that intratumoral (I.T.) injection of TT causes vascular disruption with concomitant tumor ablation in several preclinical models of cancer, in addition to various (sub)cutaneous tumors presenting in the veterinary clinic. TT has completed Phase I dose escalation trials, with some patients showing signs of abscopal effects. However, the exact molecular details underpinning its mechanism of action (MoA), together with its immunotherapeutic potential in oncology remain unclear.Methods A combination of microscopy, luciferase assays, immunofluorescence, immunoblotting, subcellular fractionation, intracellular ATP assays, phagocytosis assays and mixed lymphocyte reactions were used to probe the MoA of TT in vitro. In vivo studies with TT used MM649 xenograft, CT-26 and immune checkpoint inhibitor refractory B16-F10-OVA tumor bearing mice, the latter with or without anti-programmed cell death 1 (PD-1)/anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) mAb treatment. The effect of TT at injected and non-injected tumors was also assessed.Results Here, we show that TT induces the death of endothelial and cancer cells at therapeutically relevant concentrations via a caspase/gasdermin E-dependent pyroptopic pathway. At therapeutic doses, our data demonstrate that TT acts as a lipotoxin, binding to and promoting mitochondrial/endoplasmic reticulum (ER) dysfunction (leading to unfolded protein responsemt/ER upregulation) with subsequent ATP depletion, organelle swelling, caspase activation, gasdermin E cleavage and induction of terminal necrosis. Consistent with binding to ER membranes, we found that TT treatment promoted activation of the integrated stress response together with the release/externalization of damage-associated molecular patterns (HMGB1, ATP, calreticulin) from cancer cells in vitro and in vivo, characteristics indicative of immunogenic cell death (ICD). Confirmation of ICD in vivo was obtained through vaccination and rechallenge experiments using CT-26 colon carcinoma tumor bearing mice. Furthermore, TT also reduced tumor volume, induced immune cell infiltration, as well as improved survival in B16-F10-OVA tumor bearing mice when combined with immune checkpoint blockade.Conclusions These data demonstrate that TT is an oncolytic small molecule with multiple targets and confirms that cell death induced by this compound has the potential to augment antitumor responses to immunotherapy.https://jitc.bmj.com/content/12/4/e006602.full |
| spellingShingle | Kevin Hendrawan Kelly M Brooks Pei-Yi Yap Blake Ferguson Motoko Koyama Herlina Handoko Jenny Johns Natasa Broit Praphaporn Stewart Daniel Shelley Tracey McMahon Yi Chieh Lim Giovanni Appendino Jason K Cullen Zara C Bruce Steven M Ogbourne Paul W Reddell Glen M Boyle Peter G Parsons Jacinta L Simmons Emily S Wilson Marjorie M A de Souza Tam Hong Nguyen Alberto Pagani Victoria A Gordon Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockade Journal for ImmunoTherapy of Cancer |
| title | Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockade |
| title_full | Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockade |
| title_fullStr | Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockade |
| title_full_unstemmed | Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockade |
| title_short | Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockade |
| title_sort | tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non injected tumors to immune checkpoint blockade |
| url | https://jitc.bmj.com/content/12/4/e006602.full |
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