Pathogenesis and Therapy of Mitochondrial Diseases
type: CreativeWork
description: Mitochondria are highly dynamic organelles found in most eukaryotic cells, with a fundamental role in the generation of cellular energy through oxidative phosphorylation (OXPHOS). Critical for their function, mitochondria have retained their own genome the mitochondrial DNA, mtDNA. In mammals, replication of mtDNA is ensured by the DNA polymerase POLγ, which is composed by one catalytic subunit POLγA and two accessory subunits POLγB. Mutations in the nuclear-encoded POLG gene, coding for POLγA, are a common cause of human disease leading to a spectrum of disorders characterised by mtDNA instability, thus compromising mitochondrial function. Despite being relatively frequent, the molecular pathogenesis of POLG-related diseases is poorly understood and efficient treatments are missing, partly due to the lack of relevant in vivo models. Here, I describe the generation of two mouse models: 1) the PolgA449T/A449T mouse, which reproduces the A467T change, the most common human recessive mutation of POLG and 2) the PolgWT/Y933C mouse, which reproduces the Y955C change, the most common human dominant mutation of POLG. I focused on the use of the PolgA449T/A449T mouse and complementary in vitro techniques to provide insights into the molecular pathogenic mechanism of this POLG mutation. I describe the data showing that the mouse A449T mutation impairs DNA binding and mtDNA synthesis activities of POLγ, leading to a stalling phenotype. Most importantly, the A449T mutation also strongly impairs interaction with POLγB, the accessory subunit of the POLγ holoenzyme. This allows the free POLγA to become a substrate for LONP1 protease degradation, leading to dramatically reduced levels of POLγA in A449T mouse tissues, with consequences for the pathogenesis of the disease. In the second part of the dissertation, I explore a gene therapy approach for mitochondrial diseases associated with mutations in nuclear-encoded genes. In particular, I test the use of a novel adeno-associated vi…
abstract: Mitochondria are highly dynamic organelles found in most eukaryotic cells, with a fundamental role in the generation of cellular energy through oxidative phosphorylation (OXPHOS). Critical for their function, mitochondria have retained their own genome the mitochondrial DNA, mtDNA. In mammals, replication of mtDNA is ensured by the DNA polymerase POLγ, which is composed by one catalytic subunit POLγA and two accessory subunits POLγB. Mutations in the nuclear-encoded POLG gene, coding for POLγA, are a common cause of human disease leading to a spectrum of disorders characterised by mtDNA instability, thus compromising mitochondrial function. Despite being relatively frequent, the molecular pathogenesis of POLG-related diseases is poorly understood and efficient treatments are missing, partly due to the lack of relevant in vivo models. Here, I describe the generation of two mouse models: 1) the PolgA449T/A449T mouse, which reproduces the A467T change, the most common human recessive mutation of POLG and 2) the PolgWT/Y933C mouse, which reproduces the Y955C change, the most common human dominant mutation of POLG. I focused on the use of the PolgA449T/A449T mouse and complementary in vitro techniques to provide insights into the molecular pathogenic mechanism of this POLG mutation. I describe the data showing that the mouse A449T mutation impairs DNA binding and mtDNA synthesis activities of POLγ, leading to a stalling phenotype. Most importantly, the A449T mutation also strongly impairs interaction with POLγB, the accessory subunit of the POLγ holoenzyme. This allows the free POLγA to become a substrate for LONP1 protease degradation, leading to dramatically reduced levels of POLγA in A449T mouse tissues, with consequences for the pathogenesis of the disease. In the second part of the dissertation, I explore a gene therapy approach for mitochondrial diseases associated with mutations in nuclear-encoded genes. In particular, I test the use of a novel adeno-associated vi…
author: da Silva Pinheiro, Pedro
language: eng
keywords: mitochondria, polg, gene therapy, mitochondrial diseases
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2022-09-30
date modified: 2026-07-22T22:23:54Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.83452