Responsabile Scientifico

• OLGASI Cristina (DiMET)

Partecipanti al progetto

• OLGASI Cristina (DiMET)
• MERLIN Simone (DSS)

Titolo del progetto

• Novel cell and gene therapy approach on patient-specific endothelial cells and iPSCs-derived megakaryocytes to correct the dysfunctional release of lysosome-related organelles in Hermansky- Pudlak syndrome

Ente finanziatore

• MUR - Ministero dell'Università e Ricerca

Programma di finanziamento

• PRIN - Progetti di Ricerca di Rilevante Interesse Nazionale (dal 2020)

Bando di riferimento

• PRIN 2022 PNRR

Anno di presentazione del progetto

• 2022

Anno di approvazione del progetto

• 2023

Project ID / numero contratto

• P20222827F_001

Durata del progetto

• 24 mesi

Inizio del progetto

• 30/11/2023

Fine del progetto

• 29/11/2025

Ente capofila

• UNIUPO - Universita Degli Studi del Università del Piemonte Orientale "Amedeo Avogadro"

Altri partner del progetto

• UNIPG - Università degli Studi di Perugia

Progetto approvato - Contributo unità di ricerca del Dipartimento

• €116.309,00

Sustainable Development Goals - Agenda 2030

Keyword

• endothelial colony forming cells
• lentiviral vector
• Hermansky-Pudlak syndrome
• megakaryocytes
• induced pluripotent stem cells (iPSCs)
• cell and gene therapy

Stato del progetto

• Approvato

Abstract

• Hermansky-Pudlak syndrome (HPS) is an inherited multisystem disorder caused by a defect of lysosome-related organelles (LRO) in various tissues, like delta granules in platelets, melanosomes in melanocytes and Weibel Palade Bodies (WPB) in endothelial cells (ECs). HPS is characterized by oculocutaneous albinism with nystagmus and visual acuity loss, bleeding diathesis and, in some cases, extra-hematological complications such as pulmonary fibrosis, granulomatous colitis and immunodeficiency. HPS is caused by genetic variants in 11 different genes, all coding for proteins playing a role in the biogenesis of lysosome-related organelles. The most severe subtype of HPS is HPS-1, in which variants affect the HPS1 protein that interacts with HPS4 to compose the BLOC-3 complex. Treatment options for inherited platelet disorders (IPD), including HPS, are rather limited, unspecific, and frequently ineffective, therefore further studies are required to define the mechanism of the disease and identify an effective treatment. The mechanisms involved in the pathogenesis of HPS have already been investigated in melanosomes, alveolar epithelial cells and platelet delta granules but little is known about Weibel Palade Bodies (WPB) secretion by ECs. Recently, endothelial colony forming cells (ECFC) have been isolated from an HPS-2 patient, and in parallel with CRISPR-Cas9-engineered ECs, have been used as a cellular model to study the mechanism of maturation of WPBs and their exocytosis in HSP-2 suggesting a link between impaired endothelial cell function and bleedings. However, little is known about the maturation and storage of WPBs in HPS-1. Moreover, induced pluripotent stem cells (iPSCs) have been generated from the reprogramming of somatic cells of HSP-1 patients, but they were not differentiated in any relevant cell to be used in HSP. Therefore, this project aims to shed new light on the pathogenesis of HPS-1 and to develop for the first time a cell and gene approach to correct genetic variants causing HPS-1. Hence, we first aim to investigate the endothelial function and WPB secretion of ECFCs isolated from an HPS-1 patient. Secondly, we will generate iPSCs from an HPS-1 patient, and differentiate them into megakaryocytes and subsequently obtain platelets, to study dense granule content and release. Finally, we will develop a cell and gene therapy strategy, based on the use of lentiviral vectors carrying the HSP1 gene under the control of an endothelial or a megakaryocyte promoter, to correct the genetic defect in HPS-1 ECFCs and iPSCs-derived megakaryocytes. The findings deriving from this project will expand knowledge on a very rare inherited disorder, moreover, the concept that the bleeding phenotype of patients with HPS may be due not only to the well-known platelet defect but also to a defect in endothelial cells is very pioneering and may change the therapeutic approach to this disorder.