Modeling of monogenic diseases for physiopathological studies and pharmacological tests using specialized cells derived from iPSCs

• Grant number: 20/14635-1
• Support Opportunities: Research Projects - Thematic Grants
• Start date: December 01, 2021
• End date: November 30, 2026
• Field of knowledge: Biological Sciences - Biology
• Principal Investigator: João Bosco Pesquero
• Grantee: João Bosco Pesquero
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Pesquisadores principais: Danilo Roman Campos
• Associated researchers: Alysson Renato Muotri ; Karina Ramalho Bortoluci ; Marimélia Aparecida Porcionatto
• Associated research grant(s): 22/11408-0 - Multi-user equipment approved in grant 2020/14635-1: EVOS M7000 imaging system, AP.EMU
• Associated scholarship(s): 23/14694-6 - In vitro models for the study of the neuropathophysiology of Fabry disease, BP.PD; 24/09281-7 - Development of an Isogenic Model of Long QT Syndrome 3 using induced pluripotent human stem cells, BP.IC; 24/06444-2 - Sodium Channel Nav1.8 as a therapeutic target to treat arrhythmias in pulmonary hypertension and Fabry disease, BP.PD; + associated scholarships 23/18058-7 - Involvement of the kallikrein-kinin system in the pathophysiology of Fabry Disease from patient iPSC-derived cardiomyocytes, BP.PD; 23/00063-4 - In vitro models to study the neuropathophysiology of Fabry disease, BP.PD; 18/23367-0 - Study of the pathophysiological mechanisms of psychiatric disorders in Fabry's disease, BP.PD - associated scholarships

Abstract

Currently, one of the great challenges in medical research is to model cells in vitro capable of recapitulating physiological processes in the most similar way possible to complex organisms. The difficulty of obtaining specialized cells for studies in laboratories such as cardiomyocytes, endothelial cells and neurons has always been a challenge for the development of more accurate studies, which has been overcome in the last decade with the use of Induced Pluripotent Stem Cells (iPSCs). Since its development, iPSCs have shown their potential to differentiate into specialized cells. In addition, these cells behave like embryonic stem cells, bypassing ethical controversies, with no sample limit and with the ability to differentiate into almost all cell types. In this project we aim to use these cells in order to create an in-vitro platform for pathophysiological studies and drug analysis in 3 monogenic diseases. For this purpose, the development of cardiomyocytes and neurons derived from iPSCs with changes in genes responsible for the development of Fabry's Disease (DF) and Long QT Syndrome Type 3 (LQT3) was proposed. In addition, iPSC-derived endothelial cells carrying mutations responsible for the development of Hereditary Angioedema (HA) will also be developed. All specialized cells will be modeled from adult peripheral blood cells of patients with monogenic diseases due to or mutations induced by the CRISPr-Cas9 system. For this, iPSCs will be generated through reprogramming with episomal vectors and differentiated with specific factors. The differentiated cells will be characterized, and later analyzed in their pathophysiological processes, through molecular, fluorescent markers and electrophysiological analyzes. The use of these cells for the development of organoids may also better elucidate the biological processes involved in the accumulation of GB3 in DF, and their relationship with neural and cardiac involvement, in addition to the electrophysiological processes in LQT3 and vascular permeability in HA. The platform can also open perspectives for drug tests and bring important pharmacogenomic information capable of more assertively guiding drug prescriptions according to the type of mutation presented by the patient. (AU)

Scientific publications

(14)

(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)

RIGUETTI, MICHELLE TIVERON PASSOS; VARELA-CALAIS, PATRICIA; FERNANDES, DANILO E.; FRANCO, JOSE FRANCISCO DA SILVA; NOGUEIRA, BEATRIZ RIBEIRO; PESQUERO, JOAO B.; MASTROIANNI-KIRSZTAJN, GIANNA. Thrombomodulin Gene Mutation and Associated Predisposing Factors in Familial Collapsing Glomerulopathy. NEPHRON, v. 148, n. 7, p. 6-pg., 2024-07-01. (19/05266-5, 20/14635-1)

SOUZA, DIEGO SANTOS; MARQUES, LEISIANE PEREIRA; COSTA, ALEXANDRE DANTAS; CRUZ, JADER SANTOS; RHANA, PAULA; SANTOS-MIRANDA, ARTUR; JOVIANO-SANTOS, JULLIANE VASCONCELOS; DURCO, AIMEE OBOLARI; LINS VASCONCELOS, CARLA MARIA; ROMAN-CAMPOS, DANILO. Experimental hypothyroidism induces cardiac arrhythmias and ranolazine reverts and prevents the phenotype. Life Sciences, v. 308, p. 13-pg., 2022-09-13. (20/14635-1, 21/05584-7, 19/21304-4, 19/18918-0)

TEIXEIRA-FONSECA, JORGE LUCAS; SANTOS-MIRANDA, ARTUR; MARQUES, IVAN LOBO SOUSA; MARQUES, LEISIANE PEREIRA; ALCANTARA, FABIANA; CONCEICAO, MICHAEL RAMON DE LIMA; SOUZA, DIEGO SANTOS; GONDIM, ANTONIO NEI SANTANA; ROMAN-CAMPOS, DANILO. Eugenol delays the onset of ouabain-induced ventricular cardiac arrhythmias in guinea pigs. BASIC & CLINICAL PHARMACOLOGY & TOXICOLOGY, v. N/A, p. 11-pg., 2023-09-19. (21/15122-0, 20/14635-1, 21/05584-7, 19/18918-0)

CONCEICAO, MICHAEL RAMON DE LIMA; TEIXEIRA-FONSECA, JORGE LUCAS; ORTS, DIEGO JOSE BELATO; NASCIMENTO, DANIELLA SANTOS; DANTAS, CACIA OLIVEIRA; DE VASCONCELOS, CARLA MARIA LINS; SOUZA, DIEGO SANTOS; ROMAN-CAMPOS, DANILO. Exploring the interplay between extracellular pH and Dronedarone's pharmacological effects on cardiac function. European Journal of Pharmacology, v. 983, p. 11-pg., 2024-09-12. (24/06444-2, 22/07256-0, 22/14133-1, 21/05584-7, 20/14635-1)

CRUZ, AYEXA; GOMES, DIEGO; VERDAN, CINTIA; BRANQUINHO, JESSICA; XAVIER, MICHELE; KIRSZTAJN, GIANNA; DOS SANTOS, MAJ CALEB GUEDES MIRANDA; PESQUERO, JOAO BOSCO; CARNEIRO, ANDREIA. ACE I/D and AGT Met235Thr Polymorphisms Distinctly Affect Biomarker Levels and Risk of AKI and Exertional Rhabdomyolysis After Intense Exercise. MILITARY MEDICINE, v. N/A, p. 9-pg., 2025-02-20. (14/27198-8, 20/14635-1)

TEIXEIRA-FONSECA, JORGE LUCAS; BELATO Y ORTS, DIEGO JOSE; DA SILVA, POLYANA LEAL; CONCEICAO, MICHAEL RAMON DE LIMA; HERMES, HERNAN; PRUDENCIO, CARLOS R.; ROMAN-CAMPOS, DANILO. In Vivo Anti-Inflammatory Activity of D-Limonene in a Rat Model of Monocrotaline-Induced Pulmonary Hypertension: Implications to the. Arquivos Brasileiros de Cardiologia, v. 121, n. 12, p. 11-pg., 2024-01-01. (21/05584-7, 22/07256-0, 20/14635-1)

GUARAGNA, MARA S.; CASIMIRO, FERNANDA M. S.; VARELA, PATRICIA; DE S. FELTRAN, LUCIANA; WATANABE, ANDREIA; NEVES, PRECIL D. M. M.; PESQUERO, JOAO B.; BELANGERO, VERA M. S.; NOGUEIRA, PAULO C. K.; ONUCHIC, LUIZ F.. Past and future in vitro and in vivo approaches toward circulating factors and biomarkers in idiopathic nephrotic syndrome. Pediatric Nephrology, v. N/A, p. 17-pg., 2025-01-30. (20/02988-7, 20/15800-6, 20/14635-1)

JORGE LUCAS TEIXEIRA-FONSECA; DIEGO JOSE BELATO Y ORTS; POLYANA LEAL DA SILVA; MICHAEL RAMON DE LIMA CONCEIÇÃO; HERNAN HERMES; CARLOS R. PRUDENCIO; DANILO ROMAN-CAMPOS. Atividade Anti-Inflamatória In Vivo do D-Limoneno em um Modelo de Hipertensão Pulmonar Induzida por Monocrotalina em Ratos: Implicações à Função Cardíaca. Arquivos Brasileiros de Cardiologia, v. 121, n. 12, 2024-12-13. (20/14635-1, 22/14133-1, 21/05584-7, 22/07256-0)

SANTOS-MIRANDA, ARTUR; JOVIANO-SANTOS, V. JULLIANE; CRUZ-NASCIMENTO, TAYNARA; NERI, ELIDA ADALGISA; SOUZA, DIEGO SANTOS; MARQUES, LEISIANE PEREIRA; KRIEGER, JOSE E.; ROMAN-CAMPOS, DANILO. The fungicide Tebuconazole induces electromechanical cardiotoxicity in murine heart and human cardiomyocytes derived from induced pluripotent stem cells. Toxicology Letters, v. 359, p. 10-pg., 2022-04-15. (18/22830-9, 19/18918-0, 21/05584-7, 19/21304-4, 20/14635-1, 18/20777-3)

TEIXEIRA-FONSECA, JORGE LUCAS; SOUZA, DIEGO SANTOS; CONCEICAO, MICHAEL RAMON DE LIMA; MARQUES, LEISIANE PEREIRA; DURCO, AIMEE OBOLARI; DA SILVA, POLYANA LEAL; JOVIANO-SANTOS, JULLIANE V.; SANTOS-MIRANDA, ARTUR; ROMAN-CAMPOS, DANILO. In vivo tebuconazole administration impairs heart electrical function and facilitates the occurrence of dobutamine-induced arrhythmias: involvement of reactive oxygen species. Food and Chemical Toxicology, v. 187, p. 12-pg., 2024-04-04. (22/14133-1, 21/05584-7, 20/14635-1, 21/15122-0, 23/11912-2)

MARQUES, LEISIANE PEREIRA; SANTOS-MIRANDA, ARTUR; JOVIANO-SANTOS, JULLIANE VASCONCELOS; TEIXEIRA-FONSECA, JORGE LUCAS; ALCANTARA, FABIANA DA SILVA; SARMENTO, JAQUELINE OLIVEIRA; ROMAN-CAMPOS, DANILO. The fungicide tebuconazole modulates the sodium current of human Na_V1.5 channels expressed in HEK293 cells. Food and Chemical Toxicology, v. 180, p. 12-pg., 2023-08-30. (21/15122-0, 20/14635-1, 21/05584-7)

TEIXEIRA-FONSECA, JORGE LUCAS; JOVIANO-SANTOS, JULLIANE V.; BESERRA, SAMUEL SANTOS; CONCEICAO, MICHAEL RAMON DE LIMA; LEAL-SILVA, POLYANA; MARQUES, LEISIANE PEREIRA; SOUZA, DIEGO SANTOS; ROMAN-CAMPOS, DANILO. Exploring the involvement of TASK-1 in the control of isolated rat right atrium function from healthy animals and an experimental model of monocrotaline-induced pulmonary hypertension. NAUNYN-SCHMIEDEBERGS ARCHIVES OF PHARMACOLOGY, v. N/A, p. 14-pg., 2023-06-20. (19/21304-4, 19/18918-0, 21/15122-0, 20/14635-1, 21/05584-7)

DE CONCEICAO, MICHAEL RAMON LIMA; TEIXEIRA-FONSECA, JORGE LUCAS; MARQUES, LEISIANE PEREIRA; SOUZA, DIEGO SANTOS; ROMAN-CAMPOS, DANILO. Interaction of the antiarrhythmic drug Amiodarone with the sodium channel Nav1.5 depends on the extracellular pH. European Journal of Pharmacology, v. 960, p. 12-pg., 2023-10-24. (21/05584-7, 19/18918-0, 21/15122-0, 20/14635-1)

MARINS, MARYANA; CURIATI, MARCO ANTONIO; GOMES, CAIO PEREZ; MARTIN, RENAN PAULO; NICOLICHT-AMORIM, PRISCILA; YAMAMOTO, JOYCE UMBELINO DA SILVA; D'ALMEIDA, VANIA; MARTINS, ANA MARIA; PESQUERO, JOAO BOSCO. α-mannosidosis diagnosis in Brazilian patients with MPS-like symptoms. ORPHANET JOURNAL OF RARE DISEASES, v. 19, n. 1, p. 10-pg., 2024-11-26. (20/14635-1)