GLP Test Facility
GLP (Good Laboratory Practice) is a quality system related to the organizational process and the conditions under which non‐clinical health and environmental safety studies are planned, performed, monitored, recorded, reported and archived. A rigorous preclinical evaluation represents the foundation for the progression to clinical trials of conventional pharmaceutical products as well as Gene Therapy Medicinal Products (GTMP). GLP compliant preclinical studies ensure the quality and integrity of the data submitted to the Regulatory Agencies to support further clinical development of new therapies. Moreover, it fosters mutual acceptance of data in the international community and helps minimizing redundant testing and animal use, while maximizing protection of human health.
The GLP Test Facility at SR-Tiget is certified by the Italian Ministry of Health to perform preclinical studies of gene and cell therapies, in particular in vivo toxicity and tumorigenicity studies and analytical tests according to GLP and OECD principles. The studies performed in the GLP Test Facility have supported the approval by the regulatory bodies of clinical trials of gene therapy for beta-thalassemia (NCT02453477), MPSIH (NCT03488394), ADA-SCID and MLD (NCT01560182). In the field of gene therapy for cancer, preclinical studies by the GLP test facility have supported the approval of clinical trials in patients with acute lymphoblastic leukemia (NCT03389035) and glioblastoma (NCT03866109).
The Facility is active both for institutional projects, as well as for external customer’s projects.
Primary objectives of the Facility are to evaluate the safety of GTMP providing results to meet the regulatory standards and ensuring data and reports are generated with high standard of quality and integrity. The GLP Test Facility combines skills and expertise in gene and cell therapy research, pharmaceutical research and development, pathology and quality assurance. The complexity of the development of cell and gene therapy products is addressed by designing tailored studies to allow safety assessment of GTMP and to fulfill GLP requirements and OECD principles.
The list of studies includes:
- Toxicology and tumorigenicity studies to evaluate the potential tumorigenicity and toxicity induced by the transplantation of hematopoietic stem cells (HSC) modified by non-viral or viral vectors or by gene editing, or by direct in vivo injection of viral vectors;
- Biodistribution studies to monitor the distribution in target and off-target tissues of genetically modified human HSC, using immunodeficient murine models;
- GLP bioanalytical method validation studies to perform analysis of preclinical study samples;
- GCLP bioanalytical method validation studies to perform analysis of clinical trial samples;
- R&D studies to set up the study design and to develop analytical methods;
- R&D PATH studies for histopathological evaluation and mouse model characterization.
Specific activities include:
- Optimization according to GLP principles of protocols for HSC isolation and transduction with viral vectors, mouse conditioning, transplantation, manipulation and monitoring of clinical signs
- Development according to GLP principles of biological assays to produce and characterize the Test and Control Items
- Validation of in vitro tests, molecular and biochemical assays and cell‐based processes
- Clonogenic assays of murine and human hematopoietic progenitors
- Titration of monoclonal antibodies
- Quantitative PCR, for quantification of vector copy number (VCN) integrated in the human genome by Viia7 Real-Time PCR System Thermo Fisher Scientific
- Droplet digital PCR for quantification of VCN integrated in the human, primate and murine genome and evaluation of gene expression by QX200 Digital PCR system Bio-Rad
- Hematological analyses by hematology analyzer Procyte
- Enzymatic assays by Fluorimeter & Elisa Reader Victor – Perkin Elmer
- Flow cytometry analysis by Flow Cytometry Beckman Coulter
- In vivo studies, and histopathological evaluations are managed by the Ascentos computer system
All activities performed within GLP test facility are managed according to GLP principles by following documented steps from planning, recording, reporting and finally archiving. Experimental activities are performed following standard operating procedures (SOP) and studies are designed according to the guidelines of the International Council for Harmonization (ICH) and EMA guidelines.
Jofra Hernández R, Calabria A, Sanvito F, De Mattia F, Farinelli G, Scala S, Visigalli I, Carriglio N, De Simone M, Vezzoli M, Cecere F, Migliavacca M, Basso-Ricci L, Omrani M, Benedicenti F, Norata R, Rancoita PMV, Di Serio C, Albertini P, Cristofori P, Naldini L, Gentner B, Montini E, Aiuti A, Mortellaro A. Hematopoietic Tumors in a Mouse Model of X-linked Chronic Granulomatous Disease after Lentiviral Vector-Mediated Gene Therapy. Mol Ther. 2021 Jan 6;29(1):86-102. doi: 10.1016/j.ymthe.2020.09.030. Epub 2020 Sep 23.
Lidonnici MR, Paleari Y, Tiboni F, Mandelli G, Rossi C, Vezzoli M, Aprile A, Lederer CW, Ambrosi A, Chanut F, Sanvito F, Calabria A, Poletti V, Mavilio F, Montini E, Naldini L, Cristofori P, Ferrari G. Multiple Integrated Non-clinical Studies Predict the Safety of Lentivirus-Mediated Gene Therapy for β-Thalassemia. Mol Ther Methods Clin Dev. 2018 Sep 13;11:9-28. doi: 10.1016/j.omtm.2018.09.001. eCollection 2018 Dec 14.
Carriglio N, Klapwijk J, Hernandez RJ, Vezzoli M, Chanut F, Lowe R, Draghici E, Nord M, Albertini P, Cristofori P, Richards J, Staton H, Appleby J, Aiuti A, Sauer AV. Good Laboratory Practice Preclinical Safety Studies for GSK2696273 (MLV Vector-Based Ex Vivo Gene Therapy for Adenosine Deaminase Deficiency Severe Combined Immunodeficiency) in NSG Mice. Hum Gene Ther Clin Dev. 2017 Mar;28(1):17-27. doi: 10.1089/humc.2016.191.
Visigalli I, Delai S, Ferro F, Cecere F, Vezzoli M, Sanvito F, Chanut F, Benedicenti F, Spinozzi G, Wynn R, Calabria A, Naldini L, Montini E, Cristofori P, Biffi A. Preclinical Testing of the Safety and Tolerability of Lentiviral Vector-Mediated Above-Normal Alpha-L-Iduronidase Expression in Murine and Human Hematopoietic Cells Using Toxicology and Biodistribution Good Laboratory Practice Studies. Hum Gene Ther. 2016 Oct;27(10):813-829. doi: 10.1089/hum.2016.068.
Study Director - Validation manager
Raisa Jofra Hernandez
Vice-Archivist; SOP Administrator
Maura De Simone
GLP Pathology Manager
Marco Dante Rossini