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Development of a gene therapy model for hereditary angioedema based on SERPING1 gene edition by CRISPR-Cas9 system

Grant number: 15/25494-1
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): August 01, 2016
Effective date (End): June 30, 2019
Field of knowledge:Biological Sciences - Genetics - Human and Medical Genetics
Principal Investigator:João Bosco Pesquero
Grantee:Camila Lopes Veronez
Home Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
Associated research grant:14/27198-8 - Establishment of a center of genetic and molecular research for clinical challenges, AP.TEM
Associated scholarship(s):16/23304-3 - Angioedema induction in rats overexpressing the B2 kinin receptor: an alternative animal model for hereditary angioedema study, BE.EP.PD

Abstract

Hereditary angioedema (HAE) is a rare genetic disease resulting from a deficiency of C1 inhibitor (C1-INH) or mutations in F12 gene. In both cases, the changes lead to uncontrolled activation of the kallikrein-kinin system (KKS) culminating in excessive release of bradykinin (BK). HAE profoundly affects the quality of life, treatment is expensive and often inefficient. We propose a gene therapy model based on C1-INH gene correction (SERPING1).First of all, we intend to generate mice and cell lines with the p.A297T correspondent mutation found in Brazilian HAE families using the CRISPR-Cas9 system. Then, the mutation generated in SERPING1 will be corrected using the same system. The plasmid pSpCas9 nickase (pSpCas9n) will be used to generate C1-INH deficient C57BL/6N mice through pro-nuclear microinjections. Gene editing will be evaluated by sequencing, by SURVEYOR to check indel formation and by next generation sequencing and BLESS to check off-target events. In vitro and in vivo assays will be performed in parallel. Mice characterization will be performed by C1-INH measurements and function, as well as vascular permeability evaluation. From this point, the first tests applying an in vivo SERPING1 correction protocol using hydrodinamic injection of the constructions in the tail vein of the animal will be performed.

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
VERONEZ, CAMILA LOPES; CAMPOS, REGIS ALBUQUERQUE; CONSTANTINO-SILVA, ROSEMEIRE NAVICKAS; NICOLICHT, PRISCILA; PESQUERO, JOAO BOSCO; GRUMACH, ANETE SEVCIOVIC. Hereditary Angioedema-Associated Acute Pancreatitis in C1-Inhibitor Deficient and Normal C1-Inhibitor Patients: Case Reports and Literature Review. FRONTIERS IN MEDICINE, v. 6, APR 17 2019. Web of Science Citations: 0.
VERONEZ, CAMILA LOPES; AABOM, ANNE; MARTIN, RENAN PAULO; FILIPPELLI-SILVA, RAFAEL; GONCALVES, ROZANA FATIMA; NICOLICHT, PRISCILA; MENDES, AGATHA RIBEIRO; DA SILVA, JANE; GUILARTE, MAR; GRUMACH, ANETE SEVCIOVIC; MANSOUR, ELI; BYGUM, ANETTE; PESQUERO, JOAO BOSCO. Genetic Variation of Kallikrein-Kinin System and Related Genes in Patients With Hereditary Angioedema. FRONTIERS IN MEDICINE, v. 6, FEB 21 2019. Web of Science Citations: 0.
FELTRAN, LUCIANA S.; VARELA, PATRICIA; SILVA, ELTON DIAS; VERONEZ, CAMILA LOPES; FRANCO, MARIA CARMO; PACHECO FILHO, ALVARO; CAMARGO, MARIA FERNANDA; KOCH NOGUEIRA, PAULO CESAR; PESQUERO, JOAO BOSCO. Targeted Next-Generation Sequencing in Brazilian Children With Nephrotic Syndrome Submitted to Renal Transplant. TRANSPLANTATION, v. 101, n. 12, p. 2905-2912, DEC 2017. Web of Science Citations: 5.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.