Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy
Abstract Background Antibody-drug conjugates (ADCs) targeting the RON receptor, a tumorigenic factor contributing to cancer malignancy, has been considered as a novel strategy for cancer therapy. Here we describe a humanized antibody recognizing the RON plexin-semaphorin-integrin (PSI) domain with i...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMJ Publishing Group
2019-09-01
|
| Series: | Journal for ImmunoTherapy of Cancer |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40425-019-0732-8 |
| _version_ | 1818368088895127552 |
|---|---|
| author | Xiang-Min Tong Liang Feng Sreedhar Reddy Suthe Tian-Hao Weng Chen-Yu Hu Yi-Zhi Liu Zhi-Gang Wu Ming-Hai Wang Hang-Ping Yao |
| author_facet | Xiang-Min Tong Liang Feng Sreedhar Reddy Suthe Tian-Hao Weng Chen-Yu Hu Yi-Zhi Liu Zhi-Gang Wu Ming-Hai Wang Hang-Ping Yao |
| author_sort | Xiang-Min Tong |
| collection | DOAJ |
| description | Abstract Background Antibody-drug conjugates (ADCs) targeting the RON receptor, a tumorigenic factor contributing to cancer malignancy, has been considered as a novel strategy for cancer therapy. Here we describe a humanized antibody recognizing the RON plexin-semaphorin-integrin (PSI) domain with increased drug delivery capability for potential clinical application. Method Monoclonal antibody PCM5B14 specific to the human and monkey RON PSI domain was generated and characterized by various immunological methods. Humanized antibody H5B14 was created by grafting PCM5B14 complementarity-determining regions into human IgG1/κ acceptor frameworks and conjugated with monomethyl auristatin E and duocarmycin to form two H5B14-based ADCs. Stability of H5B14-based ADCs in human plasma was measured using hydrophobic interaction chromatography. Various biochemical and biological assays were used to determine ADC- regulated RON internalization, cell viability, spheroid formation, and death of cancer stem-like cells. Efficacies of H5B14-based ADCs in vivo were validated using tumor xenograft models. Maximal tolerated doses of H5B14-based ADCs were established in mice. Results H5B14 was highly specific to the human RON PSI domain and superior over other anti-RON ADCs in induction of RON internalization in various cancer cell lines tested. H5B14-based ADCS had a drug to antibody ratio of ~ 3.70:1 and were stable in human plasma with a minimal dissociation within a 10-day period. Functionally, H5B14-mediated drug delivery decreased cell viability at early stages with an average IC50 at ~ 20 nM in multiple cancer cell lines examined. H5B14-based ADCs also inhibited spheroid formation and caused death of cancer stem-like cells with RON+/CD44+/ESA+ phenotypes. In vivo, H5B14-based ADCs in a single injection inhibited tumor xenograft growth mediated by multiple cancer cell lines. Tumoristatic concentrations calculated from xenograft tumor models were in the range of 0.63 to 2.0 mg/kg bodyweight. Significantly, H5B14-based ADCs were capable of eradicating tumors at variable levels across multiple xenograft models regardless their malignant statuses. Toxicologically, H5B14-based ADCs were well tolerated in mice up to 60 mg/kg. Conclusion H5B14-based ADCs targeting the RON PSI domain are superior in inducing RON internalization, leading to robust drug delivery and overall inhibition and eradication of tumors in multiple xenograft models. These findings warrant H5B14-based ADCs for clinical trials in the future. |
| first_indexed | 2024-12-13T23:02:24Z |
| format | Article |
| id | doaj.art-00ae9f38d59f4b8caa9e7db02b17a3ce |
| institution | Directory Open Access Journal |
| issn | 2051-1426 |
| language | English |
| last_indexed | 2024-12-13T23:02:24Z |
| publishDate | 2019-09-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj.art-00ae9f38d59f4b8caa9e7db02b17a3ce2022-12-21T23:28:21ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262019-09-017111510.1186/s40425-019-0732-8Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapyXiang-Min Tong0Liang Feng1Sreedhar Reddy Suthe2Tian-Hao Weng3Chen-Yu Hu4Yi-Zhi Liu5Zhi-Gang Wu6Ming-Hai Wang7Hang-Ping Yao8Department of Hematology, Zhejiang Provincial People’s Hospital, Hangzhou Medical CollegeCancer Biology Research Center, Texas Tech University Health Sciences Center School of PharmacyDepartment of Pharmaceutical Sciences, Texas Tech University Health Sciences Center School of PharmacyState Key Laboratory for Diagnosis & Treatment of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of MedicineState Key Laboratory for Diagnosis & Treatment of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of MedicineState Key Laboratory for Diagnosis & Treatment of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of MedicineState Key Laboratory for Diagnosis & Treatment of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of MedicineDepartment of Hematology, Zhejiang Provincial People’s Hospital, Hangzhou Medical CollegeState Key Laboratory for Diagnosis & Treatment of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of MedicineAbstract Background Antibody-drug conjugates (ADCs) targeting the RON receptor, a tumorigenic factor contributing to cancer malignancy, has been considered as a novel strategy for cancer therapy. Here we describe a humanized antibody recognizing the RON plexin-semaphorin-integrin (PSI) domain with increased drug delivery capability for potential clinical application. Method Monoclonal antibody PCM5B14 specific to the human and monkey RON PSI domain was generated and characterized by various immunological methods. Humanized antibody H5B14 was created by grafting PCM5B14 complementarity-determining regions into human IgG1/κ acceptor frameworks and conjugated with monomethyl auristatin E and duocarmycin to form two H5B14-based ADCs. Stability of H5B14-based ADCs in human plasma was measured using hydrophobic interaction chromatography. Various biochemical and biological assays were used to determine ADC- regulated RON internalization, cell viability, spheroid formation, and death of cancer stem-like cells. Efficacies of H5B14-based ADCs in vivo were validated using tumor xenograft models. Maximal tolerated doses of H5B14-based ADCs were established in mice. Results H5B14 was highly specific to the human RON PSI domain and superior over other anti-RON ADCs in induction of RON internalization in various cancer cell lines tested. H5B14-based ADCS had a drug to antibody ratio of ~ 3.70:1 and were stable in human plasma with a minimal dissociation within a 10-day period. Functionally, H5B14-mediated drug delivery decreased cell viability at early stages with an average IC50 at ~ 20 nM in multiple cancer cell lines examined. H5B14-based ADCs also inhibited spheroid formation and caused death of cancer stem-like cells with RON+/CD44+/ESA+ phenotypes. In vivo, H5B14-based ADCs in a single injection inhibited tumor xenograft growth mediated by multiple cancer cell lines. Tumoristatic concentrations calculated from xenograft tumor models were in the range of 0.63 to 2.0 mg/kg bodyweight. Significantly, H5B14-based ADCs were capable of eradicating tumors at variable levels across multiple xenograft models regardless their malignant statuses. Toxicologically, H5B14-based ADCs were well tolerated in mice up to 60 mg/kg. Conclusion H5B14-based ADCs targeting the RON PSI domain are superior in inducing RON internalization, leading to robust drug delivery and overall inhibition and eradication of tumors in multiple xenograft models. These findings warrant H5B14-based ADCs for clinical trials in the future.http://link.springer.com/article/10.1186/s40425-019-0732-8RON receptor tyrosine kinasePSI domainMonoclonal antibodyHumanizationAntibody-drug conjugatesReceptor internalization |
| spellingShingle | Xiang-Min Tong Liang Feng Sreedhar Reddy Suthe Tian-Hao Weng Chen-Yu Hu Yi-Zhi Liu Zhi-Gang Wu Ming-Hai Wang Hang-Ping Yao Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy Journal for ImmunoTherapy of Cancer RON receptor tyrosine kinase PSI domain Monoclonal antibody Humanization Antibody-drug conjugates Receptor internalization |
| title | Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy |
| title_full | Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy |
| title_fullStr | Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy |
| title_full_unstemmed | Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy |
| title_short | Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy |
| title_sort | therapeutic efficacy of a novel humanized antibody drug conjugate recognizing plexin semaphorin integrin domain in the ron receptor for targeted cancer therapy |
| topic | RON receptor tyrosine kinase PSI domain Monoclonal antibody Humanization Antibody-drug conjugates Receptor internalization |
| url | http://link.springer.com/article/10.1186/s40425-019-0732-8 |
| work_keys_str_mv | AT xiangmintong therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy AT liangfeng therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy AT sreedharreddysuthe therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy AT tianhaoweng therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy AT chenyuhu therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy AT yizhiliu therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy AT zhigangwu therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy AT minghaiwang therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy AT hangpingyao therapeuticefficacyofanovelhumanizedantibodydrugconjugaterecognizingplexinsemaphorinintegrindomainintheronreceptorfortargetedcancertherapy |