Periodic Reporting for period 1 - PORTAL (Personalized oncology for refractory and relapsed lymphoma using humanized patient derived othotopic xenografts (ImmunePDOX))
Período documentado: 2018-04-01 hasta 2020-03-31
Lymphomas are a group of heterogenous blood cancers that arise from B or T lymphocytes and occur in 12 cases in 100,000 inhabitants each year. The most frequent B cell lymphomas are B-cell non-Hodgkin’s lymphomas (B-NHL). The 50% of B-NHL patients are refractory or relapse to the first line of treatment and 30% of them die with progressive disease. Some T cell lymphoma subtypes are also not curable and progress rapidly developing resistance to treatment. Therefore, therapeutic resistance represents a major treatment challenge for lymphoma patients. Select the best therapeutic options for each patient to overcome therapy resistance is an overriding priority. To do that it is necessary to improve our knowledge of the molecular mechanisms involved in therapy resistance and it is essential to dispose of good preclinical models for lymphoma that allow the combination of molecular and therapeutic studies. Patient derived orthotopic xenografts (PDOX) are the most advanced preclinical models in which human tumors are implanted in immunosuppressed mice at the same organ as the tumor is growing in the patient (orthotopic implantation) providing a similar tumor microenvironment. PDOX preserves the same histologic and genetic characteristics and similar pharmacological response pattern than its patient’s tumor precursor. PDOX can also be more humanized by engrafting human immune cells in the immunosuppressed mice. There is a lack of representative PDOX for lymphomas. In this project, we proposed to develop the technology to generate humanized PDOX for refractory and relapsed lymphomas to generate a proof-of-concept for their use in personalized oncology strategies in clinical practice. Our approach was to generate lymphoma PDOX by implanting patient’s tumors in lymphoid tissues and to humanize them using components of the immune system of the same patients from whom the models have been derived.
During this project we have developed the technology to generate 14 advanced PDOX for aggressive B and T lymphomas. These models are representative of different lymphoma histologies, including diffuse large B cell lymphoma, adult Burkitt lymphoma EVB negative, Hodgkin Lymphoma, Primary cutaneous large B-cell lymphoma leg type, Marginal zone B-cell lymphoma and angioimmunoblastic T-cell lymphoma. The tumor take rate obtained was 52%, as we implanted a total of 27 lymphomas. Additionally, we have generated one PDOX model of extramedullary multiple myeloma (EMM) from two EMM implanted. Growth rates (time between tumor passages), were between two to four months, with the exception of the Burkitt lymphoma model and the EMM model that showed a rapid growth, with a period between passages of three to four weeks. PDOX have been generated in NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice that are the most immunosuppressed murine models available. To humanize them with human immune system, we have tested several strategies. Initially we used peripheral blood mononuclear cells (PBMC) separated from blood samples but quantity of PBMC obtained from patients was insufficient and they generated graft versus host disease (GvsHd). Finally, we have adopted the humanization with CD8 + T lymphocytes expanded ex vivo and depleted of CD4+ T cells. Phenotypic and genetic matching between PDOX and precursor patient’s tumor were checked by histology comparison, test of surface cell markers and by genetic genotyping using six different microsatellite markers. Genetic characteristics of tumors were studied using new generation sequencing approaches (panels and/or exome sequencing) and fluorescence in situ hybridization (FISH). For the therapeutic studies, the tumors were firstly expanded in 3 to 5 animals and then reimplanted in a minimum of 8 animals for each experimental group. The experimental groups were defined together with the clinicians responsible for the patient. Only those drugs or combinations that were possible to be administered to the patient were included and adapted o experimental model to control toxicity. Results of in vivo therapeutic assays were communicated to the hematologist responsible for the patients to support therapy decisions. The project results are submitted and in preparation for publication in open access journals. Four other collaborative papers were published also in open access journals, two in Nature Communications and other two in Plos Neglected Tropical Diseases. At the end of this fellowship I was hired as principal investigator of the Catalan Institute of Oncology (ICO) / Bellvitge Biomedical Research Institute (IDIBELL) where this project has been developed and will be expanded as a new research line in the coming years. In terms of technology transfer and commercialization, a technology transfer agreement between IDIBELL and the IDIBELL Spinoff Xenopat, S.L (www.xenopat.com) is already being discussed for the commercial exploitation of the lymphoma preclinical models.
No robust methods of predicting how patients will respond to oncology drugs before treating them are used in clinical practice and drug resistance is one of the main obstacles to increase the chances of long-term survival for lymphoma patients. Our project established preclinical models to support therapy decisions with the most advanced strategies that currently exist and providing more translatable results to the patient. With this we contributed in two fundamental issues: (i) to target cancer cells as aggressively as possible from the beginning of treatment reducing unnecessary toxicity to the patient and (ii) to reduce the economic cost of patient treatment for the Health System. The development of this project allowed to initiate a new line of research in therapy resistance and orthotopic xenografts in hematologic in the ICO program ProCURE, program that has focused its studies in resistance in solid tumors and has favored starting translational research activity within the Clinical Hematology Service of ICO, which traditionally focused its scientific activity on clinical research, with the involvement of the clinicians. I will continue to lead this new research line hired as researcher of ICO/IDIBELL in the coming years. This fellowship has been a qualitative leap in my professional career. I have acquired new skills in technical aspects (as generation and humanization of patient derived preclinical models, genetic analysis, bioinformatics, among others) as well as in project management, dissemination and communication actions and scientific and industry collaboration approaches. The sum of all these aspects and experiences have had a very positive impact on my professional career and have greatly favored my re-incorporation as a principal investigator at a research center in Europe.