eRapport

Immuno-gene-therapy of prostate cancer

Prosjekt
Prosjektnummer
2011054
Ansvarlig person
Gunnar Kvalheim
Institusjon
Oslo universitetssykehus HF
Prosjektkategori
Postdoktorstipend
Helsekategori
Cancer
Forskningsaktivitet
1. Underpinning
Rapporter
2015 - sluttrapport
Immuno-gene-therapy against cancer Background/strategy: Immunotherapy in cancer patients is tasked with overcoming tumor cells that have escaped immunosurveillance. Here, we bypass tumor escape by retargeting host T cells with T cell receptors (TCRs) selected from two sources: i) Chimeric antigen-receptors (CARs) generated by combing the antigen binding region of an antibody with the intracellular regions of a TCR. ii) T cell clones from clinical responders after cancer vaccination. From long term survivors, we have identified high affinity TCRs that have arisen in vivo, recognize naturally processed antigens and mediate anti-tumor activity. Methods: TCR sequences are obtained from tumor-specific T cell clones. TCR/CAR sequences are cloned into vectors for retroviral or mRNA expression. Thereafter the TCRs/CARs are transduced or transfected to T cells. The functionality is tested in vitro and in mouse models. After establishing functional TCRs/CARs, we aim to test the constructs in phase I trials. Tumor resistance to cancer immunotherapy is investigated in patient samples, mainly with flow cytometry (in Oslo) or HLA gene analyses (Grenada). Current project status: In 2011, postdoc Jon Amund Kyte started the project by analyzing immunological data from long term survivors from our telomerase trials. He selected hTERT-specific TCRs for further studies, with regard to redirecting T-cells. He also wrote two articles reporting clinical and immunological data from the hTERT vaccine patients. Jon Amund Kyte was from July 2012 to July 2013 visiting researcher at Dr. Martin Pule’s lab, University College London. In London, Dr. Kyte cloned and characterized two hTERT-specific TCRs. Further, he started the development of a CAR against a novel cancer target. In this process, dr. Kyte learned a range of laboratory techniques that will be useful for our lab in Oslo. After returning to our Oslo lab Aug 2013, Dr. Kyte has continued to investigate the functionality of T cells redirected with the hTERT specific TCRs. He finds that both TCRs are highly potent and confer the desired specificity to transfected T cells. This work formed the basis for DOFI 14032; 2014, which in turn lead to a 9 million NOK Biotek grant from NRC for 2015-2017. The work has been presented at scientific meetings (CIMT, PIVAC) and is leading to an article that we expect to submit in 2016 (Kyte et al, manus in prep). Dr. Kyte has together with his previous MD/Master-student (forskerlinje) GL Hansen performed in depth characterisation of samples from the hTERT vaccine trials, focusing on regulatory cells countering the effect of T cell therapy and vaccination. This has lead to a paper published in 2015 (Cancer Immunology Immunotherapy 2015 Oct 26). Regarding the T-cells redirected with CARs, dr. Kyte was main supervisor for PhD-student Hilde Almåsbak. We have in 2012/2013 conducted animal studies that have produced interesting findings, suggesting that the redirected T-cells effectively and specifically kill tumor cells expressing CD19. Ms. Almåsbak’s PhD thesis was accepted June 2014. Dr. Kyte has written an article based on these data, published in 2015 (Gene Ther 2015 May; 22(5): 391-403). The project has also paved the way for a clinical trial that is under preparation in leukemia patients (protocol written by JA Kyte in 2014/2015). Dr. Kyte also has a collaborative project with Prof. Garrido's lab in Grenada, Spain. Here, the focus is to uncover mechanisms of tumor escape from immunotherapy, based on selected patients in the clinical studies that Dr. Kyte is involved in. This project has resulted in an article published Jan 2014 (del Campo, Kyte, Carretero et al Int J Canc Jan 2014). The article reports the immune escape of HLA class I-negative tumor cells and chronological sequence of appearance of tumor ß2m gene mutation in successive lesions obtained from a melanoma patient, receiving immunotherapy at our Oslo lab. The results may lead to novel cancer therapy.
2014
Main aims: 1) To develop cancer immunotherapy based on redirected T-cells 2) To analyze mechanisms making tumor cells resistant to immunotherapy 3) To bring in technology to our cancer immunotherapy center at Oslo University Hospital and establish collaboration with an international state-of-the-art laboratoryIn 2011, postdoc Jon Amund Kyte started the project by analyzing immunological data from long term survivors from our telomerase trials. He selected hTERT-specific TCRs for further studies, with regard to redirecting T-cells. He also wrote two articles reporting clinical and immunological data from the hTERT vaccine patients, both published in Clinical Cancer Research. Jon Amund Kyte was from July 2012 to July 2013 visiting researcher at Dr. Martin Pule’s lab, University College London. In London, Dr. Kyte cloned and characterized two hTERT-specific TCRs. Further, he started the development of a CAR against a novel cancer target. In this process, dr. Kyte learned a range of laboratory techniques that will be useful for our lab in Oslo. After returning to our Oslo lab Aug 2013, Dr. Kyte has continued to investigate the functionality of T cells redirected with the hTERT specific TCRs. He finds that both TCRs are highly potent and confer the desired specificity to transfected T cells. This has lead to a DOFI/patent application that was accepted by Inven2 in 2014 and formed the basis for a succesful application to Forskningsrådet for Biotek funding. Based on the hTERT TCR results, Dr. Kyte is writing an original research article that we expect to submit within 3 months (Kyte et al, manus in prep). Further, we conduct ongoing experiments aimed at bringing this concept into clinical testing. Dr. Kyte has also performed in depth characterisation of samples from the hTERT vaccine trials, focusing on regulatory cells countering the effect of T cell therapy and vaccination. Dr. Kyte has analyzed these data and expect to submit an article on this project together with his previous student (“forskerlinjestudent”) Gaute Hansen within one month. Dr. Kyte also has a collaborative project with Prof. Garrido's lab in Grenada, Spain. Here, the focus is to uncover mechanisms of tumor escape from immunotherapy, based on selected patients in the clinical studies that Dr. Kyte is involved in. This project has resulted in an article published Jan 2014 (del Campo, Kyte, Carretero et al Int J Canc Jan 2014). The article reports the immune escape of HLA class I-negative tumor cells and chronological sequence of appearance of tumor ß2m gene mutation in successive lesions obtained from a melanoma patient, receiving immunotherapy at our Oslo lab. Regarding the T-cells redirected with CARs, dr. Kyte has been main supervisor for PhD-student Hilde Almåsbak. We have in 2012/2013 conducted animal studies that have produced interesting findings, suggesting that transiently redirected T-cells effectively and specifically kill tumor cells expressing CD19. In 2013/2014, Dr. Kyte has written an article based on these data (last and corresponding author), published in Gene Therapy in 2015. Ms. Almåsbak's PhD thesis was submitted Jan 2014, and she successfully completed her PhD defence June 2014. Dr. Kyte currently works with bringing our CD19 CAR project (mRNA-mased CARs) into clinical resting. He has written a clinical protocol (testing in acute lymphocytic leukemia patinets) and works with our team at Dept. of Cell Therapy and our clinical partners (Dep. of Hematology, Rikshospitalet), in order to complete the final steps before the trial can be started. This includes preparing the documentation for clinical grade, large-scale production of CAR T cells.
2013
Main aims: 1) To develop cancer immunotherapy based on redirected T-cells 2) To analyze mechanisms making tumor cells resistant to immunotherapy 3) To bring in technology to our cancer immunotherapy center at Oslo University Hospital and establish collaboration with an international state-of-the-art laboratoryBackground: Immunotherapy in cancer patients is tasked with overcoming tumor cells that have escaped immunosurveillance. Here, we bypass tumor escape by retargeting host T cells with T cell receptors (TCRs) selected from two sources: i) Chimeric antigen-receptors (CARs) generated by combing the antigen binding region of an antibody with the intracellular regions of a TCR. ii) T cell clones from clinical responders after cancer vaccination. From long term survivors, we have identified high affinity TCRs that have arisen in vivo, recognize naturally processed antigens and mediate anti-tumor activity. Methods: TCR sequences are obtained from tumor-specific T cell clones. TCR/CAR sequences are cloned into vectors for retroviral or mRNA expression. Thereafter the TCRs/CARs are transduced or transfected to T cells. The functionality is tested in vitro and in mouse models. After establishing functional TCRs/CARs, we aim to test the constructs in phase I trials. Tumor resistance to cancer immunotherapy is investigated in patient samples, mainly with flow cytometry (in Oslo) or HLA gene analyses (Grenada). Current project status: In 2011, postdoc Jon Amund Kyte started the project by analyzing immunological data from long term survivors from our telomerase trials. He selected hTERT-specific TCRs for further studies, with regard to redirecting T-cells. He also wrote two articles reporting clinical and immunological data from the hTERT vaccine patients. Jon Amund Kyte was from July 2012 to July 2013 visiting researcher at Dr. Martin Pule’s lab, University College London. In London, Dr. Kyte cloned and characterized two hTERT-specific TCRs. Further, he started the development of a CAR against a novel cancer target. In this process, dr. Kyte learned a range of laboratory techniques that will be useful for our lab in Oslo. After returning to our Oslo lab Aug 2013, Dr. Kyte has continued to investigate the functionality of T cells redirected with the hTERT specific TCRs. He finds that both TCRs are highly potent and confer the desired specificity to transfected T cells. Dr. Kyte has also performed in depth characterisation of samples from the hTERT vaccine trials, focusing on regulatory cells countering the effect of T cell therapy and vaccination. Dr. Kyte is analyzing these data and expect to submit an article on this project together with his previous student (“forskerlinjestudent”) Gaute Hansen within a year. Regarding the T-cells redirected with CARs, dr. Kyte is main supervisor for PhD-student Hilde Almåsbak. We have in 2012/2013 conducted animal studies that have produced interesting findings, suggesting that the redirected T-cells effectively and specifically kill tumor cells expressing CD19. In 2013/2014, Dr. Kyte has written two articles based on these data (not yet published). Ms. Almåsbak’s PhD thesis has been submitted Jan 2014. Dr. Kyte also has a collaborative project with Prof. Garrido's lab in Grenada, Spain. Here, the focus is to uncover mechanisms of tumor escape from immunotherapy, based on selected patients in the clinical studies that Dr. Kyte is involved in. This project has resulted in an article published Jan 2014 (del Campo, Kyte, Carretero et al Int J Canc Jan 2014). The article reports the immune escape of HLA class I-negative tumor cells and chronological sequence of appearance of tumor ß2m gene mutation in successive lesions obtained from a melanoma patient, receiving immunotherapy at our Oslo lab.
2012
Main aims: 1) To develop cancer therapy based on redirected T-cells 2) To bring in technology to our cancer immunotherapy center at Oslo University Hospital and establish collaboration with an international state-of-the-art laboratoryBackground/strategy: Immunotherapy in cancer patients is tasked with overcoming tumor cells that have evolved in co-existence with the host immune system and escaped immunosurveillance. Here, we bypass tumor escape by retargeting host T cells with strongly cancer reactive T cell receptors (TCRs). The TCRs may be selected from two sources: i) Chimeric antigen-receptors generated by combing the antigen binding region of an antibody with the transmembrane/intracellular regions of a TCR. ii) T cell clones from long term clinical responders after vaccination: We have conducted six clinical trials with a vaccine targeting the universal tumor antigen telomerase. From long term survivors, we have identified high affinity TCRs that have arisen in vivo, recognize naturally processed antigens and mediate anti-tumor activity. Methods: TCR sequences are to be obtained from tumor-specific T cell clones. Antibodies for CAR generation are to be generated through immunization of animals with the appropriate antigen. The DNA sequences encoding TCRs/CARs will be cloned into vectors for retroviral or mRNA expression. Thereafter the TCRs/CARs are to be retrovirally transduced into T cells or transfected as mRNA for transient expression. The functionality is to be tested in vitro and in mouse models. After establishing functional TCRs/CARs, we plan a phase I/II trial. Current project status: In 2011, postdoc Jon Amund Kyte started the project by analyzing immunological data from long term survivors from our telomerase trials. He selected hTERT-specific TCRs for further studies, with regard to redirecting T-cells. He also wrote an article together with Dr. Brunsvig, reporting clinical and immunological data from these long term survivors (Clin Cancer Res. 2011 Nov 1;17(21):6847-57). Regarding the T-cells redirected with CARs, dr. Kyte is main supervisor for PhD-student Hilde Almåsbak. We have in 2012 conducted animal studies that have produced interesting findings, suggesting that the redirected T-cells effectively and specifically kill tumor cells expressing CD19. The results are expected to be published in 2013 and have paved the way for a clinical trial that is under preparation. Jon Amund Kyte has from July 2012 started his period as visiting researcher at Dr. Martin Pule’s lab, University College London. Kyte has to date cloned and characterized tumor-specific TCRs. Further, he has started the development of a CAR against a novel cancer target. In this process, dr. Kyte is learning a range of laboratory techniques that will be useful for our lab in Oslo upon his return. Through Dr. Kyte’s work in London we aim at starting long term collaborative projects with dr. Martin Pule’s group. Dr. Kyte has to this aim had detailed discussions with Dr. Pule. We plan to apply for 1-2 technicians/PhD-students working at these joint projects, co-funded by us and University College London.
2011
Main aims: 1) To develop cancer therapy based on redirected T-cells 2) To bring in technology from an international state-of-the-art laboratory to our cancer immunotherapy center at Oslo University HospitalBackground/strategy: Immunotherapy in cancer patients is tasked with overcoming tumor cells that have evolved in co-existence with the host immune system and escaped immunosurveillance. Here, we bypass tumor escape by retargeting host T cells with strongly cancer reactive T cell receptors (TCRs) from other individuals. The TCRs may be selected from three sources: i) Allo-reactive TCRs against tumor-associated antigens, generated and selected ex vivo ii) Chimeric antigen-receptors generated by combing the antigen binding region of an antibody with the transmembrane/intracellular regions of a TCR. iii) T cell clones from long term clinical responders after vaccination: We have conducted six clinical trials with a vaccine targeting the universal tumor antigen telomerase. From long term survivors, we have identified high affinity TCRs that have arisen in vivo, recognize naturally processed antigens and mediate anti-tumor activity. Methods: The TCR/CAR sequences will be retrovirally transduced into T cells or transfected as mRNA for transient expression. The functionality is to be tested in vitro and in mouse models. After establishing functional TCRs/CARs, we plan a phase I/II trial. Current project status: Postdoc Jon Amund Kyte started the project in 2011 by analyzing immunological data from long term survivors from our telomerase trials. He selected hTERT-specific TCRs for further studies, with regard to redirecting T-cells. He also wrote an article together with Dr. Brunsvig, reporting clinical and immunological data from these long term survivors (Clin Cancer Res. 2011 Nov 1;17(21):6847-57). Regarding the T-cells redirected with CARs, dr. Kyte is main supervisor for PhD-student Hilde Almåsbak, who has conducted in vitro experiments with a CD19-specific CAR and started in vivo animal studies late 2011. The results so far suggest that the redirected T-cells effectively and specifically kill tumor cells expressing CD19. The start of the postdoc project was delayed for about 6 months due to a change in time point for Dr. Kyte's research period at an international laboratory. Dr. Kyte has used these 6 months to complete his specialisation in oncology. We have also decided that he will work at Dr. Pule's laboratory in London, rather than at Dr. Brenner's lab in Houston. Their two laboratories are closely associated (see below). These changes have been reported to Health Region South East. Plans: Dr. Kyte is working with the development of redirected T-cells. He has established collaboration with several experts at Oslo University Hospital, Dr. Malcolm Brenner, Houston, and Dr. Martin Pule, University College London . To develop the project further, Dr. Kyte plans to work for about a year at Dr. Pule's laboratory in London. Dr. Pule’s group is associated with Dr. Brenner’s institute in Houston, which is vastly experienced within the field of redirected T-cells, including translation into the clinic. Through this collaboration, we aim to bring in state-of-the-art technology to Oslo and to initiate the first clinical studies with redirected T-cells in Norway.
Vitenskapelige artikler
Hansen GL, Gaudernack G, Brunsvig PF, Cvancarova M, Kyte JA

Immunological factors influencing clinical outcome in lung cancer patients after telomerase peptide vaccination.

Cancer Immunol Immunother 2015 Dec;64(12):1609-21. Epub 2015 okt 26

PMID: 26498005

Almåsbak H, Walseng E, Kristian A, Myhre MR, Suso EM, Munthe LA, Andersen JT, Wang MY, Kvalheim G, Gaudernack G, Kyte JA

Inclusion of an IgG1-Fc spacer abrogates efficacy of CD19 CAR T cells in a xenograft mouse model.

Gene Ther 2015 May;22(5):391-403. Epub 2015 feb 5

PMID: 25652098

del Campo Ana B, Kyte Jon Amund, Carretero Javier, Zinchencko Svitlana, Méndez Rosa, González-Aseguinolaza Gloria, Ruiz-Cabello Francisco, Aamdal Steinar, Gaudernack Gustav, Garrido Federico, Aptsiauri Natalia

Immune escape of cancer cells with beta2-microglobulin loss over the course of metastatic melanoma.

Int J Cancer 2014 Jan 1;134(1):102-13. Epub 2013 jul 16

PMID: 23784959

Brunsvig Paal Fr, Kyte Jon Amund, Kersten Christian, Sundstrøm Stein, Møller Mona, Nyakas Marta, Hansen Gaute L, Gaudernack Gustav, Aamdal Steinar

Telomerase peptide vaccination in NSCLC: a phase II trial in stage III patients vaccinated after chemoradiotherapy and an 8-year update on a phase I/II trial.

Clin Cancer Res 2011 Nov;17(21):6847-57. Epub 2011 sep 14

PMID: 21918169

Doktorgrader
Hilde Almåsbak

Adoptive T cell therapy for B-cell malignancies: Transient redirection of T cells with chimeric antigen receptor targeting CD19

Disputert:
juni 2014
Hovedveileder:
Jon Amund Kyte
Deltagere
  • Jon Amund Kyte Postdoktorstipendiat

eRapport er utarbeidet av Sølvi Lerfald og Reidar Thorstensen, Regionalt kompetansesenter for klinisk forskning, Helse Vest RHF, og videreutvikles av de fire RHF-ene i fellesskap, med støtte fra Helse Vest IKT

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