Mitochondrial Genetics

Lead Research Organisation: University of Cambridge

Abstract

In eukaryotic organisms almost all genetic information is encoded in DNA present in the nucleus of the cell, but a small DNA molecule inhabits mitochondria, cellular structures that provide energy from food for the cells to use. Mitochondrial DNA contains genes that are vital for the physiological functioning of the cell. Defects that directly or indirectly affect mitochondrial genes cause human diseases. We are still do not know how mitochondrial genes work exactly. One of the ways to investigate the role of a gene, or to discover its biological function, it to change or disrupt DNA, and then to look for the effect on cultured cells, or on the whole organism. These methods of genetic modification are often powerful ways of studying disease genes encoded in the nucleus, but they cannot be applied to mammalian mitochondrial DNA. Therefore, our research goals are to identify new genes regulating mitochondria, define how these mitochondrial genes operate and to provide the technology to allow mammalian mitochondrial DNA to be modified genetically. It could be an invaluable way of understanding mitochondrial diseases and for advancing the quest for therapies.

Technical Summary

Human mitochondria have their own genome (mtDNA) that is two hundred thousand times smaller that the nuclear genome (nDNA) and encodes only thirteen essential subunits of the oxidative phosphorylation (OXPHOS) machinery. Given the relatively small size of the mitochondrial genome our detailed understanding of the molecular mechanism that govern mitochondrial gene maintenance and expression is still surprisingly sketchy. Defects of mtDNA and nDNA are well-recognised cause of genetic disorders that have diverse clinical manifestations ranging from progressive muscle weakness to fatal infantile disease. Much of our understanding of mitochondrial molecular genetics has come from studying rare mitochondrial disorders. Currently there are no effective treatments for these diseases and there are very few animal models for them. The slow progress in gathering more data on how mitochondrial genome is regulated is, in large part, owing to our current inability to edit mtDNA. Genome engineering techniques, analogous to the ones routinely used to modify the nuclear DNA, are not in hand, so we are unable to interrogate the role of cis-acting elements for RNA expression as well as transcription or replication. Furthermore, we are still unable to introduce pathogenic mtDNA mutations at will so their effect could be studied in animal models. In particular, the programme focuses on: (i) Studying the basic mechanisms of mitochondrial genome regulation with the main focus on the novel proteins responsible for nuclolytic processing of precursor mitochondrial RNA, polyadenylation of mitochondrial messenger RNA, post-transcriptional nucleotide modification of mitochondrial RNA and mitoribosome biogenesis. (ii) The analysis of the key aspects of mitochondrial genome regulation in samples derived from patients affected with mitochondrial disease. This analysis is a source of valuable insights into the pathomechanisms of human disease, and also into basic mitochondrial molecular genetics. Importantly. They provide patients with a molecular diagnosis for prevention (prenatal genetic diagnosis) and counselling. (iii) Developing new approaches for the genetic modification of mitochondria of living cells. The key long-term aim is to establish routine methods of genetic modification of mammalian mitochondria. Enzymatic methods are being developed to modify the mitochondrial genome e.g. engineered zinc finger nucleases are delivered to mitochondria in order to deplete cells selectively of specific mtDNA variants.

Publications

10 25 50
 
Description Diagnosis of mitochondrial disease
Geographic Reach Multiple continents/international 
Policy Influence Type Citation in clinical reviews
Impact improvements in clinical service delivery via e.g. pre-natal diagnosis
 
Description Pre-clinical approach to treatment of mtDNA disease
Geographic Reach Multiple continents/international 
Policy Influence Type Citation in clinical reviews
 
Description Biochemical Society General Travel Grant
Amount £450 (GBP)
Organisation Biochemical Society 
Sector Learned Society
Country United Kingdom
Start 06/2017 
End 06/2017
 
Description Boehringer Ingelheim Fonds Travel Grant
Amount € 1,060 (EUR)
Organisation Boehringer Ingelheim 
Department Boehringer Ingelheim Fonds
Sector Charity/Non Profit
Country Germany
Start 04/2017 
End 04/2017
 
Description Homerton College Research Grant
Amount £400 (GBP)
Organisation University of Cambridge 
Sector Academic/University
Country United Kingdom
Start 06/2017 
End 06/2017
 
Description REMIX ITN
Amount € 3,920,000 (EUR)
Organisation Marie Sklodowska-Curie Actions 
Sector Academic/University
Country Global
Start 10/2017 
End 10/2020
 
Description The Champ Foundation Grant
Amount $100,000 (USD)
Organisation The Champ Foundation 
Start 05/2017 
End 05/2019
 
Title In vivo mtDNA manipulation using programmable nucleases 
Description Over the past decade mitochondrial DNA (mtDNA) has transitioned to broader relevance across diverse fields in biology and medicine. This has largely been thanks to the recognition of mitochondria as a major biochemical hub, the discovery of mitochondrial dysfunction in various diseases and a number of high-profile attempts at preventing hereditary mitochondrial disease using three-parent in vitro fertilisation, or mitochondrial replacement therapy (1-4). Mitochondrial diseases are a genetically diverse group of hereditary, multi-system disorders that are incurable and largely untreatable, with heterogeneous penetrance, presentation and prognosis. The majority of mitochondrial diseases are transmitted through maternally inherited mutations in mtDNA, and patients suffering with these disorders present a substantial disease burden (5). Mitochondrial DNA is a small, circular genome, encoding 13 essential protein subunits of the respiratory chain complexes and ATP synthase. As the mitochondrial genome is multi-copy, with anything from 100-10,000 copies per cell depending on tissue type, mutated mtDNA co-exists with healthy wild-type mtDNA in most patients suffering from mitochondrial disease, a phenomenon known as heteroplasmy. Patient symptoms and outcomes are closely correlated with the extent of mutation load, and approaches to treatment of mtDNA disease by shifting the heteroplasmic ratio in favour of wild-type mtDNA have long been desired and pursued. As mtDNA engineering lags many decades behind its nuclear counterpart, owing to various technical challenges (6) that likely negate the possibility of a functional CRISPR/Cas9 platform in mammalian mitochondria (7), it has not been possible to produce animal models of mtDNA disease until recently. The m.5024C>T tRNAALA strain is the only available mouse model of heteroplasmic mtDNA disease, bearing a pathogenic point mutation that results in biochemical hallmarks of mtDNA disease (diminished steady-state levels of the affected mt-tRNAALA) at high levels of mutant heteroplasmy in cardiac tissue (8). The mouse m.5024C>T mutation is equivalent to the human m.5591G>A mutation in mt-tRNAALA, associated with an adult-onset mitochondrial disease (9). This animal model is also, more broadly, clinically relevant to a large number of mtDNA diseases caused by mutations in mitochondrial tRNAs. In past work from our laboratory, we have engineered a new class of programmable, mitochondria-specific nuclease: the mitochondrially-targeted zinc finger-nuclease (mtZFN), which we have demonstrated can specifically eliminate mutated mtDNA in patient-derived cell models, resulting in recovery of associated cellular dysfunction (10-11). Aspects of this work are protected by the MRC patent on mtZFNs (US9139628 B2). In our recent work we achieved the successful translation of mtZFN technology from in vitro studies to the disease-relevant m.5024C>T tRNAALA mouse model. Through the experiments presented in the manuscript, we demonstrate that: 1) mtZFNs can be engineered to selectively eliminate m.5024C>T mutation with single nucleotide specificity. 2) When delivered systemically by adeno-associated virus (AAV), mtZFNs are capable of robustly manipulating mtDNA heteroplasmy of the targeted tissue. 3) Treatment-induced shifts in mtDNA heteroplasmy are accompanied by recovery of molecular and biochemical phenotypes in cardiac tissue of the m.5024C>T tRNAALA mouse model. 4) We conclude that mtZFNs are an effective tool for the treatment of heteroplasmic mitochondrial disease in mammalian animal models, with potential for development as a new class of therapy. References: [1] Craven L, et al. 2010. Nature. 465: 82-5. [2] Tachibana M, et al. 2013. Nature. 493: 627-31. [3] Hyslop LA, et al. 2016. Nature. 534: 383-6. [4] Kang E, et al. 2016. Nature. 540: 270-275. [5] Gorman GS, et al. 2015. Ann. Neurol. 77: 753-9. [6] Patananan AN, et al. 2016. Cell Metab. 23: 785-96. [7] Gammage PA, et al., 2018. Trends. Genet. in press [8] Kauppila JHK, et al., 2016. Cell Rep. 16: 2980-90. [9] Swalwell H, et al. 2006. Neurology. 66: 447-9. [10] Gammage PA, et al. 2014. EMBO Mol. Med. 6: 458-66. [11] Gammage PA, et al. 2016. Nucleic Acids Res. 44: 7804-16. 
Type Of Material Technology assay or reagent 
Year Produced 2017 
Provided To Others? No  
Impact Taken together, this study constitute proof-of-principle that a potential cure for all heteroplasmic mitochondrial disease using mtZFNs. We describe possibly universal therapeutic for heteroplasmic mtDNA disease represent a potential transformation in the future prospects of mitochondrial disease, bringing the promise of a cure to an area of medicine that lacks efficacious treatments, let alone curative therapy. Additionally, the existence of viable therapeutics, such as mtZFN, could alter the balance of ethical arguments surrounding the use mitochondrial replacement therapy techniques (1-4), which are controversial and have been licensed partially on the basis of the desperate state of mitochondrial medicine at present. 
URL http://www.mrc-mbu.cam.ac.uk/projects/2308/genetic-modification-mitochondrial-genome
 
Title MPAT-Seq: Next-generation sequencing-based analysis of RNA polyadenylation 
Description MPAT-Seq approach allowes to perform the transcriptome-wide parallel assessment of 3' ends of mitochondrial tRNAs. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact The further application of this technique may be useful to determine the regulation of the 3' end RNA metabolism in other studies 
 
Title Quantitative gradient fractionation mass spectrometry (qGFMS) 
Description This method is based on SILAC proteomics and sucrose density gradient separation of mitoribosomal subunits 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Pending 
 
Title mTUNE 
Description We developed cell model with defined levels of m.8993T>G mutation heteroplasmy, mTUNE, to investigate the metabolic underpinnings of mitochondrial dysfunction 
Type Of Material Cell line 
Year Produced 2018 
Provided To Others? Yes  
Impact We uncovered a novel link between glycolysis and mitochondrial dysfunction mediated by reductive carboxylation of glutamine. 
 
Title mTUNE cell system 
Description Cell lines that have been through the mTUNE process resulting in heteroplasmy of 22%, 54%, 89% and a cell line containing 1% wild-type mtDNA. The remaining mtDNA all these cell lines contain the mutation m.8993T>G. - Cells containing 22%, 54% and 89% wild type mtDNA are isogenic deriving from a 143B human osteosarcoma (HOS) cell line provided by Columbia University. - Cells containing 1% wild type mtDNA are derived from the 143B HOS cell line, but is clonally different than 22%, 54% and 89% wild type mtDNA. The generation and analysis of the cell lines are described in the following publications: Gammage PA, Gaude E, Van Haute L, Rebelo-Guiomar P, Jackson CB, Rorbach J, Pekalski ML, Robinson AJ, Charpentier M, Concordet JP, Frezza C, Minczuk M. Nucleic Acids Res. 2016 Sep 19;44(16):7804-16. doi: 10.1093/nar/gkw676. Epub 2016 Jul 27 Gaude E, Schmidt C, Gammage PA, Dugourd A, Blacker T, Chew SP, Saez-Rodriguez J, O'Neill JS, Szabadkai G, Minczuk M, Frezza C. Mol Cell. 2018 Feb 15;69(4):581-593.e7. doi:10.1016/j.molcel.2018.01.034. 
Type Of Material Data analysis technique 
Year Produced 2018 
Provided To Others? Yes  
Impact This model allows for studying effects of mitochondrial dysfunction 
 
Description Analysis of a novel nuclease in patients presenting combined OXPHOS deficiencies. 
Organisation Columbia University
Department College of Physicians & Surgeons
Country United States 
Sector Academic/University 
PI Contribution In 2010 we identified a novel mitochondrial deoxyribonuclease. In order to understand its function in human mtDNA maintenance we have studied the phenotypes of gene inactivation by RNAi and overexpression and analysed the biochemical activity of the recombinant enzyme. In 2011 we started a collaboration with five other groups as they had identified a mutation in the coding gene in six individuals from three unrelated families presenting combined OXPHOS deficiency. Currently, we are confirming the phenotypes observed in the RNAi experiments using patient-derived fibroblasts and performing complementation studies. New patients identified in 2016
Collaborator Contribution following the family clinical history and provided clinical data identified mutations
Impact The project is multidisciplinary as it involves collaboration between neurologists and molecular biologists. Two papers published: Loss-of-function mutations in MGME1 impair mtDNA replication and cause multisystemic mitochondrial disease. Kornblum C, Nicholls TJ, Haack TB, Schöler S, Peeva V, Danhauser K, Hallmann K, Zsurka G, Rorbach J, Iuso A, Wieland T, Sciacco M, Ronchi D, Comi GP, Moggio M, Quinzii CM, DiMauro S, Calvo SE, Mootha VK, Klopstock T, Strom TM, Meitinger T, Minczuk M, Kunz WS, Prokisch H. Nat Genet. 2013 Feb;45(2):214-9. doi: 10.1038/ng.2501. Linear mtDNA fragments and unusual mtDNA rearrangements associated with pathological deficiency of MGME1 exonuclease. Nicholls TJ, Zsurka G, Peeva V, Schöler S, Szczesny RJ, Cysewski D, Reyes A, Kornblum C, Sciacco M, Moggio M, Dziembowski A, Kunz WS, Minczuk M. Hum Mol Genet. 2014 Dec 1;23(23):6147-62. doi: 10.1093/hmg/ddu336. We identified a new disease gene which is now routinely screened in patients with mitochondrial disease
Start Year 2011
 
Description Analysis of a novel nuclease in patients presenting combined OXPHOS deficiencies. 
Organisation Harvard University
Department Harvard Medical School
Country United States 
Sector Academic/University 
PI Contribution In 2010 we identified a novel mitochondrial deoxyribonuclease. In order to understand its function in human mtDNA maintenance we have studied the phenotypes of gene inactivation by RNAi and overexpression and analysed the biochemical activity of the recombinant enzyme. In 2011 we started a collaboration with five other groups as they had identified a mutation in the coding gene in six individuals from three unrelated families presenting combined OXPHOS deficiency. Currently, we are confirming the phenotypes observed in the RNAi experiments using patient-derived fibroblasts and performing complementation studies. New patients identified in 2016
Collaborator Contribution following the family clinical history and provided clinical data identified mutations
Impact The project is multidisciplinary as it involves collaboration between neurologists and molecular biologists. Two papers published: Loss-of-function mutations in MGME1 impair mtDNA replication and cause multisystemic mitochondrial disease. Kornblum C, Nicholls TJ, Haack TB, Schöler S, Peeva V, Danhauser K, Hallmann K, Zsurka G, Rorbach J, Iuso A, Wieland T, Sciacco M, Ronchi D, Comi GP, Moggio M, Quinzii CM, DiMauro S, Calvo SE, Mootha VK, Klopstock T, Strom TM, Meitinger T, Minczuk M, Kunz WS, Prokisch H. Nat Genet. 2013 Feb;45(2):214-9. doi: 10.1038/ng.2501. Linear mtDNA fragments and unusual mtDNA rearrangements associated with pathological deficiency of MGME1 exonuclease. Nicholls TJ, Zsurka G, Peeva V, Schöler S, Szczesny RJ, Cysewski D, Reyes A, Kornblum C, Sciacco M, Moggio M, Dziembowski A, Kunz WS, Minczuk M. Hum Mol Genet. 2014 Dec 1;23(23):6147-62. doi: 10.1093/hmg/ddu336. We identified a new disease gene which is now routinely screened in patients with mitochondrial disease
Start Year 2011
 
Description Analysis of a novel nuclease in patients presenting combined OXPHOS deficiencies. 
Organisation Ludwig Maximilian University of Munich (LMU Munich)
Department Department of Human Genetics
Country Germany 
Sector Academic/University 
PI Contribution In 2010 we identified a novel mitochondrial deoxyribonuclease. In order to understand its function in human mtDNA maintenance we have studied the phenotypes of gene inactivation by RNAi and overexpression and analysed the biochemical activity of the recombinant enzyme. In 2011 we started a collaboration with five other groups as they had identified a mutation in the coding gene in six individuals from three unrelated families presenting combined OXPHOS deficiency. Currently, we are confirming the phenotypes observed in the RNAi experiments using patient-derived fibroblasts and performing complementation studies. New patients identified in 2016
Collaborator Contribution following the family clinical history and provided clinical data identified mutations
Impact The project is multidisciplinary as it involves collaboration between neurologists and molecular biologists. Two papers published: Loss-of-function mutations in MGME1 impair mtDNA replication and cause multisystemic mitochondrial disease. Kornblum C, Nicholls TJ, Haack TB, Schöler S, Peeva V, Danhauser K, Hallmann K, Zsurka G, Rorbach J, Iuso A, Wieland T, Sciacco M, Ronchi D, Comi GP, Moggio M, Quinzii CM, DiMauro S, Calvo SE, Mootha VK, Klopstock T, Strom TM, Meitinger T, Minczuk M, Kunz WS, Prokisch H. Nat Genet. 2013 Feb;45(2):214-9. doi: 10.1038/ng.2501. Linear mtDNA fragments and unusual mtDNA rearrangements associated with pathological deficiency of MGME1 exonuclease. Nicholls TJ, Zsurka G, Peeva V, Schöler S, Szczesny RJ, Cysewski D, Reyes A, Kornblum C, Sciacco M, Moggio M, Dziembowski A, Kunz WS, Minczuk M. Hum Mol Genet. 2014 Dec 1;23(23):6147-62. doi: 10.1093/hmg/ddu336. We identified a new disease gene which is now routinely screened in patients with mitochondrial disease
Start Year 2011
 
Description Analysis of a novel nuclease in patients presenting combined OXPHOS deficiencies. 
Organisation Oslo University Hospital
Department Department of Medical Genetics
Country Norway 
Sector Hospitals 
PI Contribution In 2010 we identified a novel mitochondrial deoxyribonuclease. In order to understand its function in human mtDNA maintenance we have studied the phenotypes of gene inactivation by RNAi and overexpression and analysed the biochemical activity of the recombinant enzyme. In 2011 we started a collaboration with five other groups as they had identified a mutation in the coding gene in six individuals from three unrelated families presenting combined OXPHOS deficiency. Currently, we are confirming the phenotypes observed in the RNAi experiments using patient-derived fibroblasts and performing complementation studies. New patients identified in 2016
Collaborator Contribution following the family clinical history and provided clinical data identified mutations
Impact The project is multidisciplinary as it involves collaboration between neurologists and molecular biologists. Two papers published: Loss-of-function mutations in MGME1 impair mtDNA replication and cause multisystemic mitochondrial disease. Kornblum C, Nicholls TJ, Haack TB, Schöler S, Peeva V, Danhauser K, Hallmann K, Zsurka G, Rorbach J, Iuso A, Wieland T, Sciacco M, Ronchi D, Comi GP, Moggio M, Quinzii CM, DiMauro S, Calvo SE, Mootha VK, Klopstock T, Strom TM, Meitinger T, Minczuk M, Kunz WS, Prokisch H. Nat Genet. 2013 Feb;45(2):214-9. doi: 10.1038/ng.2501. Linear mtDNA fragments and unusual mtDNA rearrangements associated with pathological deficiency of MGME1 exonuclease. Nicholls TJ, Zsurka G, Peeva V, Schöler S, Szczesny RJ, Cysewski D, Reyes A, Kornblum C, Sciacco M, Moggio M, Dziembowski A, Kunz WS, Minczuk M. Hum Mol Genet. 2014 Dec 1;23(23):6147-62. doi: 10.1093/hmg/ddu336. We identified a new disease gene which is now routinely screened in patients with mitochondrial disease
Start Year 2011
 
Description Analysis of a novel nuclease in patients presenting combined OXPHOS deficiencies. 
Organisation University Hospital Bonn
Department Department of Epileptology
Country Germany 
Sector Hospitals 
PI Contribution In 2010 we identified a novel mitochondrial deoxyribonuclease. In order to understand its function in human mtDNA maintenance we have studied the phenotypes of gene inactivation by RNAi and overexpression and analysed the biochemical activity of the recombinant enzyme. In 2011 we started a collaboration with five other groups as they had identified a mutation in the coding gene in six individuals from three unrelated families presenting combined OXPHOS deficiency. Currently, we are confirming the phenotypes observed in the RNAi experiments using patient-derived fibroblasts and performing complementation studies. New patients identified in 2016
Collaborator Contribution following the family clinical history and provided clinical data identified mutations
Impact The project is multidisciplinary as it involves collaboration between neurologists and molecular biologists. Two papers published: Loss-of-function mutations in MGME1 impair mtDNA replication and cause multisystemic mitochondrial disease. Kornblum C, Nicholls TJ, Haack TB, Schöler S, Peeva V, Danhauser K, Hallmann K, Zsurka G, Rorbach J, Iuso A, Wieland T, Sciacco M, Ronchi D, Comi GP, Moggio M, Quinzii CM, DiMauro S, Calvo SE, Mootha VK, Klopstock T, Strom TM, Meitinger T, Minczuk M, Kunz WS, Prokisch H. Nat Genet. 2013 Feb;45(2):214-9. doi: 10.1038/ng.2501. Linear mtDNA fragments and unusual mtDNA rearrangements associated with pathological deficiency of MGME1 exonuclease. Nicholls TJ, Zsurka G, Peeva V, Schöler S, Szczesny RJ, Cysewski D, Reyes A, Kornblum C, Sciacco M, Moggio M, Dziembowski A, Kunz WS, Minczuk M. Hum Mol Genet. 2014 Dec 1;23(23):6147-62. doi: 10.1093/hmg/ddu336. We identified a new disease gene which is now routinely screened in patients with mitochondrial disease
Start Year 2011
 
Description Analysis of a novel nuclease in patients presenting combined OXPHOS deficiencies. 
Organisation University of Milan
Department Department of Neurology
Country Italy 
Sector Academic/University 
PI Contribution In 2010 we identified a novel mitochondrial deoxyribonuclease. In order to understand its function in human mtDNA maintenance we have studied the phenotypes of gene inactivation by RNAi and overexpression and analysed the biochemical activity of the recombinant enzyme. In 2011 we started a collaboration with five other groups as they had identified a mutation in the coding gene in six individuals from three unrelated families presenting combined OXPHOS deficiency. Currently, we are confirming the phenotypes observed in the RNAi experiments using patient-derived fibroblasts and performing complementation studies. New patients identified in 2016
Collaborator Contribution following the family clinical history and provided clinical data identified mutations
Impact The project is multidisciplinary as it involves collaboration between neurologists and molecular biologists. Two papers published: Loss-of-function mutations in MGME1 impair mtDNA replication and cause multisystemic mitochondrial disease. Kornblum C, Nicholls TJ, Haack TB, Schöler S, Peeva V, Danhauser K, Hallmann K, Zsurka G, Rorbach J, Iuso A, Wieland T, Sciacco M, Ronchi D, Comi GP, Moggio M, Quinzii CM, DiMauro S, Calvo SE, Mootha VK, Klopstock T, Strom TM, Meitinger T, Minczuk M, Kunz WS, Prokisch H. Nat Genet. 2013 Feb;45(2):214-9. doi: 10.1038/ng.2501. Linear mtDNA fragments and unusual mtDNA rearrangements associated with pathological deficiency of MGME1 exonuclease. Nicholls TJ, Zsurka G, Peeva V, Schöler S, Szczesny RJ, Cysewski D, Reyes A, Kornblum C, Sciacco M, Moggio M, Dziembowski A, Kunz WS, Minczuk M. Hum Mol Genet. 2014 Dec 1;23(23):6147-62. doi: 10.1093/hmg/ddu336. We identified a new disease gene which is now routinely screened in patients with mitochondrial disease
Start Year 2011
 
Description Designing mitochondrial zinc finger nucleases 
Organisation Sangamo Biosciences, Inc
Country United States 
Sector Private 
PI Contribution Developing mitochondrially-targeted engineered nucleases to eliminate human mtDNA molecules with pathogenic point mutations and large-scale deletions
Collaborator Contribution Assembling and validating zinc finger proteins
Impact Several papers e.g. Minczuk, M., Kolasinska-Zwierz, P., Murphy, M.P., Papworth, M.A. (2010) Construction and testing of engineered zinc-finger proteins for sequence-specific modification of mtDNA. Nat Protoc 5, 342-56 Gammage, P.A., Rorbach, J., Vincent, A.I., Rebar, E.J, Minczuk, M. (2014) Mitochondrially-targeted ZFNs for selective degradation of pathogenic mitochondrial genomes bearing large-scale deletions or point mutations. EMBO Mol Med 6, 458-466 In this paper we are reporting an improved version of mitochondrially-targeted engineered zinc-finger nucleases (mtZFNs) designed to eliminate human mtDNA molecules with pathogenic point mutations and large-scale deletions. Expression of mtZFNs successfully led to a reduction in the mutant mtDNA haplotype load, and subsequent repopulation of wildtype mtDNA was found to restore mitochondrial respiratory function in a cybrid cell model. This research was highlighted by: Moraes C. T. (2014) A magic bullet to specifically eliminate mutated mitochondrial genomes from patients' cells. EMBO Mol. Med. 6, 434-435 Gammage, P.A., Van Haute, L., Minczuk, M. (2015) Engineered mtZFNs for manipulation of human mitochondrial DNA heteroplasmy Methods Mol. Biol. Mitochondrial DNA McKenzie, M (Ed)
Start Year 2009
 
Description Manipulating mtDNA heteroplasmy in vivo using engineered nucleases 
Organisation Max Planck Society
Department Max Planck Institute for the Biology of Ageing
Country Germany 
Sector Academic/University 
PI Contribution We have generated a mtZFN library specific to mtDNA mutation present in a model strain, tested them in mouse embryonic fibroblasts (MEF) from the mt-tRNAAla mouse line and identified constructs capable of effective reduction of mutant load. We will deliver these ZFNs affected mice with optimal mtZFN constructs by organtropic, recombinant adeno-associated viruses (AAVs). We will also manipulte mtDNA heteroplasmy in germline using mtZFN and mitoTALENs (the latter obtained in collaboration with Uni of Miami)
Collaborator Contribution Providing mouse model, help and expertise in mtZFN research that may led to generation of pre-clinical data useful for a development of an effective treatment of mtDNA disease in the future.
Impact pending, likely to be societal. Some protocols for generation of mtZFN published: Gammage PA, Van Haute L, Minczuk M. Engineered mtZFNs for Manipulation of Human Mitochondrial DNA Heteroplasmy. Methods Mol Biol. 2016;1351:145-62. doi: 10.1007/978-1-4939-3040-1_11 Vectors available at Addgen
Start Year 2015
 
Description Manipulating mtDNA heteroplasmy in vivo using engineered nucleases 
Organisation University of Miami
Country United States 
Sector Academic/University 
PI Contribution We have generated a mtZFN library specific to mtDNA mutation present in a model strain, tested them in mouse embryonic fibroblasts (MEF) from the mt-tRNAAla mouse line and identified constructs capable of effective reduction of mutant load. We will deliver these ZFNs affected mice with optimal mtZFN constructs by organtropic, recombinant adeno-associated viruses (AAVs). We will also manipulte mtDNA heteroplasmy in germline using mtZFN and mitoTALENs (the latter obtained in collaboration with Uni of Miami)
Collaborator Contribution Providing mouse model, help and expertise in mtZFN research that may led to generation of pre-clinical data useful for a development of an effective treatment of mtDNA disease in the future.
Impact pending, likely to be societal. Some protocols for generation of mtZFN published: Gammage PA, Van Haute L, Minczuk M. Engineered mtZFNs for Manipulation of Human Mitochondrial DNA Heteroplasmy. Methods Mol Biol. 2016;1351:145-62. doi: 10.1007/978-1-4939-3040-1_11 Vectors available at Addgen
Start Year 2015
 
Description Mitochondria in drug toxicity 
Organisation AstraZeneca
Department Research and Development AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution Hosting and training a collaborative post-doc
Collaborator Contribution Providing access to a high-throughput measurements of mitochondrial function.
Impact ongoing
Start Year 2014
 
Description Mitochondrial RNA metabolism and human disease 
Organisation Carlo Besta Neurological Institute
Country Italy 
Sector Public 
PI Contribution We analyse molecular phenotypes associated with mutations in muclaer genes involved in mitochondrial RNA processing and post-transcriptional modification.
Collaborator Contribution Identification of mutations by next-generation exome sequencing in patients with combined OXPHOS defects
Impact Several papers e.g: Van Haute L, Dietmann S, Kremer L, Hussain S, Pearce SF, Powell CA, Rorbach J, Lantaff R, Blanco S, Sauer S, Kotzaeridou U, Hoffmann GF, Memari Y, Kolb-Kokocinski A, Durbin R, Mayr JA, Frye M, Prokisch H, Minczuk M.Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016 Jun 30;7:12039. doi: 10.1038/ncomms12039. Haack, T.B.*, Kopajtich, R.*, Freisinger, P.*, Wieland, T., Rorbach, J, Nicholls, T.J., Enrico Baruffini, E., Walther, A., Danhauser, K., Zimmermann, F.A., Husain, R.A., Schum, J., Mundy, H., Ferrero, I., Strom, T.M., Meitinger, T., Taylor, R.W., Minczuk, M**., Mayr, J.A., Prokisch, H.** (2013) ELAC2 Mutations Cause a Mitochondrial RNA Processing Defect Associated with Hypertrophic Cardiomyopathy. Am J Hum Genet 93, 211-223 Kopajtich, R.,* Nicholls, T.J.,* Rorbach, J.,* Freisinger, P., Mandel, H., Vanlander, A., Ghezzi, D., Carrozzo, R., Taylor, R.W., Marquard, K., Murayama, K., Wieland, T., Schwarzmayr, T., Mayr, J.A., Pearce, S. F., Powell, C. Saada, A., Ohtake, A., Invernizzi, F., Lamantea, E., Sommerville, E. W., Pyle, A., Chinnery, P. F., Crushell, E., Okazaki, Y., Kohda, M., Kishita, Y., Tokuzawa, Y., Smet, J., Régal, L., Lorber, A., Khoury, A., Zeviani, M., Strom, T. M., Meitinger, T., Bertini, E. S., Van Coster, R., Klopstock, T., Haack, T. B., Minczuk, M.,** Prokisch, H.** (-) Mutations in GTPBP3 cause a mitochondrial translation defect associated with hypertrophic cardiomyopathy, lactic acidosis and encephalopathy Am J Hum Genet 95, 708-20 Vanlander, A.V., Menten, B., Smet, J., De Meirleir, L., Sante, T., De Paepe, B., Seneca, S., Pearce, S.F., Powell, C.A., Vergult, S., Michotte, A., De Latter, E., Vantomme, L., Minczuk, M. and Van Coster, R. (-) Two siblings with homozygous pathogenic splice site mutation in mitochondrial asparaginyl-tRNA synthetase (NARS2) Hum. Mutat Mutat 36, 222-231 Powell, C.A.*, Kopajtich, R.*, D'Souza, A.R., Rorbach, J., Dallabona, C., Donnini, C., Alston, C.L., Griffin, H., Pyle, A., Chinnery, P.F., Strom, T.M., Meitinger, T., Rodenburg, R.J., Schottmann, G., Schuelke, M., Romain, N., Haller, R., Ferrero, I., Haack, T.B., Taylor, R.W., Prokisch, H.**, Minczuk, M. (2015) Mutations in TRMT5 cause a defect in post-transcriptional modification of mitochondrial tRNA associated with multiple respiratory-chain deficiencies. Am J Hum Genet. 97,319-328 Coughlin, C.R. Scharer, G.H., Friederich, M.W., Yu, H.C., Geiger, E.A., Creadon-Swindell, G., Collins, A.E., Vanlander, A.V., Coster, R.V., Powell, C.A., Swanson, M.A., Minczuk, M., Van Hove, J.L., Shaikh, T.H. (2015) Mutations in the mitochondrial cysteinyl-tRNA synthase gene, CARS2, lead to a severe epileptic encephalopathy and complex movement disorder. J Med Genet. 52,532-540 and more
Start Year 2011
 
Description Mitochondrial RNA metabolism and human disease 
Organisation Columbia University Medical Center
Country United States 
Sector Academic/University 
PI Contribution We analyse molecular phenotypes associated with mutations in muclaer genes involved in mitochondrial RNA processing and post-transcriptional modification.
Collaborator Contribution Identification of mutations by next-generation exome sequencing in patients with combined OXPHOS defects
Impact Several papers e.g: Van Haute L, Dietmann S, Kremer L, Hussain S, Pearce SF, Powell CA, Rorbach J, Lantaff R, Blanco S, Sauer S, Kotzaeridou U, Hoffmann GF, Memari Y, Kolb-Kokocinski A, Durbin R, Mayr JA, Frye M, Prokisch H, Minczuk M.Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016 Jun 30;7:12039. doi: 10.1038/ncomms12039. Haack, T.B.*, Kopajtich, R.*, Freisinger, P.*, Wieland, T., Rorbach, J, Nicholls, T.J., Enrico Baruffini, E., Walther, A., Danhauser, K., Zimmermann, F.A., Husain, R.A., Schum, J., Mundy, H., Ferrero, I., Strom, T.M., Meitinger, T., Taylor, R.W., Minczuk, M**., Mayr, J.A., Prokisch, H.** (2013) ELAC2 Mutations Cause a Mitochondrial RNA Processing Defect Associated with Hypertrophic Cardiomyopathy. Am J Hum Genet 93, 211-223 Kopajtich, R.,* Nicholls, T.J.,* Rorbach, J.,* Freisinger, P., Mandel, H., Vanlander, A., Ghezzi, D., Carrozzo, R., Taylor, R.W., Marquard, K., Murayama, K., Wieland, T., Schwarzmayr, T., Mayr, J.A., Pearce, S. F., Powell, C. Saada, A., Ohtake, A., Invernizzi, F., Lamantea, E., Sommerville, E. W., Pyle, A., Chinnery, P. F., Crushell, E., Okazaki, Y., Kohda, M., Kishita, Y., Tokuzawa, Y., Smet, J., Régal, L., Lorber, A., Khoury, A., Zeviani, M., Strom, T. M., Meitinger, T., Bertini, E. S., Van Coster, R., Klopstock, T., Haack, T. B., Minczuk, M.,** Prokisch, H.** (-) Mutations in GTPBP3 cause a mitochondrial translation defect associated with hypertrophic cardiomyopathy, lactic acidosis and encephalopathy Am J Hum Genet 95, 708-20 Vanlander, A.V., Menten, B., Smet, J., De Meirleir, L., Sante, T., De Paepe, B., Seneca, S., Pearce, S.F., Powell, C.A., Vergult, S., Michotte, A., De Latter, E., Vantomme, L., Minczuk, M. and Van Coster, R. (-) Two siblings with homozygous pathogenic splice site mutation in mitochondrial asparaginyl-tRNA synthetase (NARS2) Hum. Mutat Mutat 36, 222-231 Powell, C.A.*, Kopajtich, R.*, D'Souza, A.R., Rorbach, J., Dallabona, C., Donnini, C., Alston, C.L., Griffin, H., Pyle, A., Chinnery, P.F., Strom, T.M., Meitinger, T., Rodenburg, R.J., Schottmann, G., Schuelke, M., Romain, N., Haller, R., Ferrero, I., Haack, T.B., Taylor, R.W., Prokisch, H.**, Minczuk, M. (2015) Mutations in TRMT5 cause a defect in post-transcriptional modification of mitochondrial tRNA associated with multiple respiratory-chain deficiencies. Am J Hum Genet. 97,319-328 Coughlin, C.R. Scharer, G.H., Friederich, M.W., Yu, H.C., Geiger, E.A., Creadon-Swindell, G., Collins, A.E., Vanlander, A.V., Coster, R.V., Powell, C.A., Swanson, M.A., Minczuk, M., Van Hove, J.L., Shaikh, T.H. (2015) Mutations in the mitochondrial cysteinyl-tRNA synthase gene, CARS2, lead to a severe epileptic encephalopathy and complex movement disorder. J Med Genet. 52,532-540 and more
Start Year 2011
 
Description Mitochondrial RNA metabolism and human disease 
Organisation Helmholtz Zentrum München
Department Institute of Human Genetics
Country Germany 
Sector Public 
PI Contribution We analyse molecular phenotypes associated with mutations in muclaer genes involved in mitochondrial RNA processing and post-transcriptional modification.
Collaborator Contribution Identification of mutations by next-generation exome sequencing in patients with combined OXPHOS defects
Impact Several papers e.g: Van Haute L, Dietmann S, Kremer L, Hussain S, Pearce SF, Powell CA, Rorbach J, Lantaff R, Blanco S, Sauer S, Kotzaeridou U, Hoffmann GF, Memari Y, Kolb-Kokocinski A, Durbin R, Mayr JA, Frye M, Prokisch H, Minczuk M.Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016 Jun 30;7:12039. doi: 10.1038/ncomms12039. Haack, T.B.*, Kopajtich, R.*, Freisinger, P.*, Wieland, T., Rorbach, J, Nicholls, T.J., Enrico Baruffini, E., Walther, A., Danhauser, K., Zimmermann, F.A., Husain, R.A., Schum, J., Mundy, H., Ferrero, I., Strom, T.M., Meitinger, T., Taylor, R.W., Minczuk, M**., Mayr, J.A., Prokisch, H.** (2013) ELAC2 Mutations Cause a Mitochondrial RNA Processing Defect Associated with Hypertrophic Cardiomyopathy. Am J Hum Genet 93, 211-223 Kopajtich, R.,* Nicholls, T.J.,* Rorbach, J.,* Freisinger, P., Mandel, H., Vanlander, A., Ghezzi, D., Carrozzo, R., Taylor, R.W., Marquard, K., Murayama, K., Wieland, T., Schwarzmayr, T., Mayr, J.A., Pearce, S. F., Powell, C. Saada, A., Ohtake, A., Invernizzi, F., Lamantea, E., Sommerville, E. W., Pyle, A., Chinnery, P. F., Crushell, E., Okazaki, Y., Kohda, M., Kishita, Y., Tokuzawa, Y., Smet, J., Régal, L., Lorber, A., Khoury, A., Zeviani, M., Strom, T. M., Meitinger, T., Bertini, E. S., Van Coster, R., Klopstock, T., Haack, T. B., Minczuk, M.,** Prokisch, H.** (-) Mutations in GTPBP3 cause a mitochondrial translation defect associated with hypertrophic cardiomyopathy, lactic acidosis and encephalopathy Am J Hum Genet 95, 708-20 Vanlander, A.V., Menten, B., Smet, J., De Meirleir, L., Sante, T., De Paepe, B., Seneca, S., Pearce, S.F., Powell, C.A., Vergult, S., Michotte, A., De Latter, E., Vantomme, L., Minczuk, M. and Van Coster, R. (-) Two siblings with homozygous pathogenic splice site mutation in mitochondrial asparaginyl-tRNA synthetase (NARS2) Hum. Mutat Mutat 36, 222-231 Powell, C.A.*, Kopajtich, R.*, D'Souza, A.R., Rorbach, J., Dallabona, C., Donnini, C., Alston, C.L., Griffin, H., Pyle, A., Chinnery, P.F., Strom, T.M., Meitinger, T., Rodenburg, R.J., Schottmann, G., Schuelke, M., Romain, N., Haller, R., Ferrero, I., Haack, T.B., Taylor, R.W., Prokisch, H.**, Minczuk, M. (2015) Mutations in TRMT5 cause a defect in post-transcriptional modification of mitochondrial tRNA associated with multiple respiratory-chain deficiencies. Am J Hum Genet. 97,319-328 Coughlin, C.R. Scharer, G.H., Friederich, M.W., Yu, H.C., Geiger, E.A., Creadon-Swindell, G., Collins, A.E., Vanlander, A.V., Coster, R.V., Powell, C.A., Swanson, M.A., Minczuk, M., Van Hove, J.L., Shaikh, T.H. (2015) Mutations in the mitochondrial cysteinyl-tRNA synthase gene, CARS2, lead to a severe epileptic encephalopathy and complex movement disorder. J Med Genet. 52,532-540 and more
Start Year 2011
 
Description Mitochondrial RNA metabolism and human disease 
Organisation Newcastle University
Department School of Biomedical Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution We analyse molecular phenotypes associated with mutations in muclaer genes involved in mitochondrial RNA processing and post-transcriptional modification.
Collaborator Contribution Identification of mutations by next-generation exome sequencing in patients with combined OXPHOS defects
Impact Several papers e.g: Van Haute L, Dietmann S, Kremer L, Hussain S, Pearce SF, Powell CA, Rorbach J, Lantaff R, Blanco S, Sauer S, Kotzaeridou U, Hoffmann GF, Memari Y, Kolb-Kokocinski A, Durbin R, Mayr JA, Frye M, Prokisch H, Minczuk M.Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016 Jun 30;7:12039. doi: 10.1038/ncomms12039. Haack, T.B.*, Kopajtich, R.*, Freisinger, P.*, Wieland, T., Rorbach, J, Nicholls, T.J., Enrico Baruffini, E., Walther, A., Danhauser, K., Zimmermann, F.A., Husain, R.A., Schum, J., Mundy, H., Ferrero, I., Strom, T.M., Meitinger, T., Taylor, R.W., Minczuk, M**., Mayr, J.A., Prokisch, H.** (2013) ELAC2 Mutations Cause a Mitochondrial RNA Processing Defect Associated with Hypertrophic Cardiomyopathy. Am J Hum Genet 93, 211-223 Kopajtich, R.,* Nicholls, T.J.,* Rorbach, J.,* Freisinger, P., Mandel, H., Vanlander, A., Ghezzi, D., Carrozzo, R., Taylor, R.W., Marquard, K., Murayama, K., Wieland, T., Schwarzmayr, T., Mayr, J.A., Pearce, S. F., Powell, C. Saada, A., Ohtake, A., Invernizzi, F., Lamantea, E., Sommerville, E. W., Pyle, A., Chinnery, P. F., Crushell, E., Okazaki, Y., Kohda, M., Kishita, Y., Tokuzawa, Y., Smet, J., Régal, L., Lorber, A., Khoury, A., Zeviani, M., Strom, T. M., Meitinger, T., Bertini, E. S., Van Coster, R., Klopstock, T., Haack, T. B., Minczuk, M.,** Prokisch, H.** (-) Mutations in GTPBP3 cause a mitochondrial translation defect associated with hypertrophic cardiomyopathy, lactic acidosis and encephalopathy Am J Hum Genet 95, 708-20 Vanlander, A.V., Menten, B., Smet, J., De Meirleir, L., Sante, T., De Paepe, B., Seneca, S., Pearce, S.F., Powell, C.A., Vergult, S., Michotte, A., De Latter, E., Vantomme, L., Minczuk, M. and Van Coster, R. (-) Two siblings with homozygous pathogenic splice site mutation in mitochondrial asparaginyl-tRNA synthetase (NARS2) Hum. Mutat Mutat 36, 222-231 Powell, C.A.*, Kopajtich, R.*, D'Souza, A.R., Rorbach, J., Dallabona, C., Donnini, C., Alston, C.L., Griffin, H., Pyle, A., Chinnery, P.F., Strom, T.M., Meitinger, T., Rodenburg, R.J., Schottmann, G., Schuelke, M., Romain, N., Haller, R., Ferrero, I., Haack, T.B., Taylor, R.W., Prokisch, H.**, Minczuk, M. (2015) Mutations in TRMT5 cause a defect in post-transcriptional modification of mitochondrial tRNA associated with multiple respiratory-chain deficiencies. Am J Hum Genet. 97,319-328 Coughlin, C.R. Scharer, G.H., Friederich, M.W., Yu, H.C., Geiger, E.A., Creadon-Swindell, G., Collins, A.E., Vanlander, A.V., Coster, R.V., Powell, C.A., Swanson, M.A., Minczuk, M., Van Hove, J.L., Shaikh, T.H. (2015) Mutations in the mitochondrial cysteinyl-tRNA synthase gene, CARS2, lead to a severe epileptic encephalopathy and complex movement disorder. J Med Genet. 52,532-540 and more
Start Year 2011
 
Description Mitochondrial RNA metabolism and human disease 
Organisation Radboud University Nijmegen
Country Netherlands 
Sector Academic/University 
PI Contribution We analyse molecular phenotypes associated with mutations in muclaer genes involved in mitochondrial RNA processing and post-transcriptional modification.
Collaborator Contribution Identification of mutations by next-generation exome sequencing in patients with combined OXPHOS defects
Impact Several papers e.g: Van Haute L, Dietmann S, Kremer L, Hussain S, Pearce SF, Powell CA, Rorbach J, Lantaff R, Blanco S, Sauer S, Kotzaeridou U, Hoffmann GF, Memari Y, Kolb-Kokocinski A, Durbin R, Mayr JA, Frye M, Prokisch H, Minczuk M.Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016 Jun 30;7:12039. doi: 10.1038/ncomms12039. Haack, T.B.*, Kopajtich, R.*, Freisinger, P.*, Wieland, T., Rorbach, J, Nicholls, T.J., Enrico Baruffini, E., Walther, A., Danhauser, K., Zimmermann, F.A., Husain, R.A., Schum, J., Mundy, H., Ferrero, I., Strom, T.M., Meitinger, T., Taylor, R.W., Minczuk, M**., Mayr, J.A., Prokisch, H.** (2013) ELAC2 Mutations Cause a Mitochondrial RNA Processing Defect Associated with Hypertrophic Cardiomyopathy. Am J Hum Genet 93, 211-223 Kopajtich, R.,* Nicholls, T.J.,* Rorbach, J.,* Freisinger, P., Mandel, H., Vanlander, A., Ghezzi, D., Carrozzo, R., Taylor, R.W., Marquard, K., Murayama, K., Wieland, T., Schwarzmayr, T., Mayr, J.A., Pearce, S. F., Powell, C. Saada, A., Ohtake, A., Invernizzi, F., Lamantea, E., Sommerville, E. W., Pyle, A., Chinnery, P. F., Crushell, E., Okazaki, Y., Kohda, M., Kishita, Y., Tokuzawa, Y., Smet, J., Régal, L., Lorber, A., Khoury, A., Zeviani, M., Strom, T. M., Meitinger, T., Bertini, E. S., Van Coster, R., Klopstock, T., Haack, T. B., Minczuk, M.,** Prokisch, H.** (-) Mutations in GTPBP3 cause a mitochondrial translation defect associated with hypertrophic cardiomyopathy, lactic acidosis and encephalopathy Am J Hum Genet 95, 708-20 Vanlander, A.V., Menten, B., Smet, J., De Meirleir, L., Sante, T., De Paepe, B., Seneca, S., Pearce, S.F., Powell, C.A., Vergult, S., Michotte, A., De Latter, E., Vantomme, L., Minczuk, M. and Van Coster, R. (-) Two siblings with homozygous pathogenic splice site mutation in mitochondrial asparaginyl-tRNA synthetase (NARS2) Hum. Mutat Mutat 36, 222-231 Powell, C.A.*, Kopajtich, R.*, D'Souza, A.R., Rorbach, J., Dallabona, C., Donnini, C., Alston, C.L., Griffin, H., Pyle, A., Chinnery, P.F., Strom, T.M., Meitinger, T., Rodenburg, R.J., Schottmann, G., Schuelke, M., Romain, N., Haller, R., Ferrero, I., Haack, T.B., Taylor, R.W., Prokisch, H.**, Minczuk, M. (2015) Mutations in TRMT5 cause a defect in post-transcriptional modification of mitochondrial tRNA associated with multiple respiratory-chain deficiencies. Am J Hum Genet. 97,319-328 Coughlin, C.R. Scharer, G.H., Friederich, M.W., Yu, H.C., Geiger, E.A., Creadon-Swindell, G., Collins, A.E., Vanlander, A.V., Coster, R.V., Powell, C.A., Swanson, M.A., Minczuk, M., Van Hove, J.L., Shaikh, T.H. (2015) Mutations in the mitochondrial cysteinyl-tRNA synthase gene, CARS2, lead to a severe epileptic encephalopathy and complex movement disorder. J Med Genet. 52,532-540 and more
Start Year 2011
 
Description Mitochondrial RNA metabolism and human disease 
Organisation South Australian Clinical Genetics Service
Country Australia 
Sector Hospitals 
PI Contribution We analyse molecular phenotypes associated with mutations in muclaer genes involved in mitochondrial RNA processing and post-transcriptional modification.
Collaborator Contribution Identification of mutations by next-generation exome sequencing in patients with combined OXPHOS defects
Impact Several papers e.g: Van Haute L, Dietmann S, Kremer L, Hussain S, Pearce SF, Powell CA, Rorbach J, Lantaff R, Blanco S, Sauer S, Kotzaeridou U, Hoffmann GF, Memari Y, Kolb-Kokocinski A, Durbin R, Mayr JA, Frye M, Prokisch H, Minczuk M.Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016 Jun 30;7:12039. doi: 10.1038/ncomms12039. Haack, T.B.*, Kopajtich, R.*, Freisinger, P.*, Wieland, T., Rorbach, J, Nicholls, T.J., Enrico Baruffini, E., Walther, A., Danhauser, K., Zimmermann, F.A., Husain, R.A., Schum, J., Mundy, H., Ferrero, I., Strom, T.M., Meitinger, T., Taylor, R.W., Minczuk, M**., Mayr, J.A., Prokisch, H.** (2013) ELAC2 Mutations Cause a Mitochondrial RNA Processing Defect Associated with Hypertrophic Cardiomyopathy. Am J Hum Genet 93, 211-223 Kopajtich, R.,* Nicholls, T.J.,* Rorbach, J.,* Freisinger, P., Mandel, H., Vanlander, A., Ghezzi, D., Carrozzo, R., Taylor, R.W., Marquard, K., Murayama, K., Wieland, T., Schwarzmayr, T., Mayr, J.A., Pearce, S. F., Powell, C. Saada, A., Ohtake, A., Invernizzi, F., Lamantea, E., Sommerville, E. W., Pyle, A., Chinnery, P. F., Crushell, E., Okazaki, Y., Kohda, M., Kishita, Y., Tokuzawa, Y., Smet, J., Régal, L., Lorber, A., Khoury, A., Zeviani, M., Strom, T. M., Meitinger, T., Bertini, E. S., Van Coster, R., Klopstock, T., Haack, T. B., Minczuk, M.,** Prokisch, H.** (-) Mutations in GTPBP3 cause a mitochondrial translation defect associated with hypertrophic cardiomyopathy, lactic acidosis and encephalopathy Am J Hum Genet 95, 708-20 Vanlander, A.V., Menten, B., Smet, J., De Meirleir, L., Sante, T., De Paepe, B., Seneca, S., Pearce, S.F., Powell, C.A., Vergult, S., Michotte, A., De Latter, E., Vantomme, L., Minczuk, M. and Van Coster, R. (-) Two siblings with homozygous pathogenic splice site mutation in mitochondrial asparaginyl-tRNA synthetase (NARS2) Hum. Mutat Mutat 36, 222-231 Powell, C.A.*, Kopajtich, R.*, D'Souza, A.R., Rorbach, J., Dallabona, C., Donnini, C., Alston, C.L., Griffin, H., Pyle, A., Chinnery, P.F., Strom, T.M., Meitinger, T., Rodenburg, R.J., Schottmann, G., Schuelke, M., Romain, N., Haller, R., Ferrero, I., Haack, T.B., Taylor, R.W., Prokisch, H.**, Minczuk, M. (2015) Mutations in TRMT5 cause a defect in post-transcriptional modification of mitochondrial tRNA associated with multiple respiratory-chain deficiencies. Am J Hum Genet. 97,319-328 Coughlin, C.R. Scharer, G.H., Friederich, M.W., Yu, H.C., Geiger, E.A., Creadon-Swindell, G., Collins, A.E., Vanlander, A.V., Coster, R.V., Powell, C.A., Swanson, M.A., Minczuk, M., Van Hove, J.L., Shaikh, T.H. (2015) Mutations in the mitochondrial cysteinyl-tRNA synthase gene, CARS2, lead to a severe epileptic encephalopathy and complex movement disorder. J Med Genet. 52,532-540 and more
Start Year 2011
 
Description Mitochondrial RNA metabolism and human disease 
Organisation University College London
Department Faculty of Medical Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution We analyse molecular phenotypes associated with mutations in muclaer genes involved in mitochondrial RNA processing and post-transcriptional modification.
Collaborator Contribution Identification of mutations by next-generation exome sequencing in patients with combined OXPHOS defects
Impact Several papers e.g: Van Haute L, Dietmann S, Kremer L, Hussain S, Pearce SF, Powell CA, Rorbach J, Lantaff R, Blanco S, Sauer S, Kotzaeridou U, Hoffmann GF, Memari Y, Kolb-Kokocinski A, Durbin R, Mayr JA, Frye M, Prokisch H, Minczuk M.Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016 Jun 30;7:12039. doi: 10.1038/ncomms12039. Haack, T.B.*, Kopajtich, R.*, Freisinger, P.*, Wieland, T., Rorbach, J, Nicholls, T.J., Enrico Baruffini, E., Walther, A., Danhauser, K., Zimmermann, F.A., Husain, R.A., Schum, J., Mundy, H., Ferrero, I., Strom, T.M., Meitinger, T., Taylor, R.W., Minczuk, M**., Mayr, J.A., Prokisch, H.** (2013) ELAC2 Mutations Cause a Mitochondrial RNA Processing Defect Associated with Hypertrophic Cardiomyopathy. Am J Hum Genet 93, 211-223 Kopajtich, R.,* Nicholls, T.J.,* Rorbach, J.,* Freisinger, P., Mandel, H., Vanlander, A., Ghezzi, D., Carrozzo, R., Taylor, R.W., Marquard, K., Murayama, K., Wieland, T., Schwarzmayr, T., Mayr, J.A., Pearce, S. F., Powell, C. Saada, A., Ohtake, A., Invernizzi, F., Lamantea, E., Sommerville, E. W., Pyle, A., Chinnery, P. F., Crushell, E., Okazaki, Y., Kohda, M., Kishita, Y., Tokuzawa, Y., Smet, J., Régal, L., Lorber, A., Khoury, A., Zeviani, M., Strom, T. M., Meitinger, T., Bertini, E. S., Van Coster, R., Klopstock, T., Haack, T. B., Minczuk, M.,** Prokisch, H.** (-) Mutations in GTPBP3 cause a mitochondrial translation defect associated with hypertrophic cardiomyopathy, lactic acidosis and encephalopathy Am J Hum Genet 95, 708-20 Vanlander, A.V., Menten, B., Smet, J., De Meirleir, L., Sante, T., De Paepe, B., Seneca, S., Pearce, S.F., Powell, C.A., Vergult, S., Michotte, A., De Latter, E., Vantomme, L., Minczuk, M. and Van Coster, R. (-) Two siblings with homozygous pathogenic splice site mutation in mitochondrial asparaginyl-tRNA synthetase (NARS2) Hum. Mutat Mutat 36, 222-231 Powell, C.A.*, Kopajtich, R.*, D'Souza, A.R., Rorbach, J., Dallabona, C., Donnini, C., Alston, C.L., Griffin, H., Pyle, A., Chinnery, P.F., Strom, T.M., Meitinger, T., Rodenburg, R.J., Schottmann, G., Schuelke, M., Romain, N., Haller, R., Ferrero, I., Haack, T.B., Taylor, R.W., Prokisch, H.**, Minczuk, M. (2015) Mutations in TRMT5 cause a defect in post-transcriptional modification of mitochondrial tRNA associated with multiple respiratory-chain deficiencies. Am J Hum Genet. 97,319-328 Coughlin, C.R. Scharer, G.H., Friederich, M.W., Yu, H.C., Geiger, E.A., Creadon-Swindell, G., Collins, A.E., Vanlander, A.V., Coster, R.V., Powell, C.A., Swanson, M.A., Minczuk, M., Van Hove, J.L., Shaikh, T.H. (2015) Mutations in the mitochondrial cysteinyl-tRNA synthase gene, CARS2, lead to a severe epileptic encephalopathy and complex movement disorder. J Med Genet. 52,532-540 and more
Start Year 2011
 
Description Mitochondrial RNA metabolism and human disease 
Organisation University of Ghent
Country Belgium 
Sector Academic/University 
PI Contribution We analyse molecular phenotypes associated with mutations in muclaer genes involved in mitochondrial RNA processing and post-transcriptional modification.
Collaborator Contribution Identification of mutations by next-generation exome sequencing in patients with combined OXPHOS defects
Impact Several papers e.g: Van Haute L, Dietmann S, Kremer L, Hussain S, Pearce SF, Powell CA, Rorbach J, Lantaff R, Blanco S, Sauer S, Kotzaeridou U, Hoffmann GF, Memari Y, Kolb-Kokocinski A, Durbin R, Mayr JA, Frye M, Prokisch H, Minczuk M.Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3. Nat Commun. 2016 Jun 30;7:12039. doi: 10.1038/ncomms12039. Haack, T.B.*, Kopajtich, R.*, Freisinger, P.*, Wieland, T., Rorbach, J, Nicholls, T.J., Enrico Baruffini, E., Walther, A., Danhauser, K., Zimmermann, F.A., Husain, R.A., Schum, J., Mundy, H., Ferrero, I., Strom, T.M., Meitinger, T., Taylor, R.W., Minczuk, M**., Mayr, J.A., Prokisch, H.** (2013) ELAC2 Mutations Cause a Mitochondrial RNA Processing Defect Associated with Hypertrophic Cardiomyopathy. Am J Hum Genet 93, 211-223 Kopajtich, R.,* Nicholls, T.J.,* Rorbach, J.,* Freisinger, P., Mandel, H., Vanlander, A., Ghezzi, D., Carrozzo, R., Taylor, R.W., Marquard, K., Murayama, K., Wieland, T., Schwarzmayr, T., Mayr, J.A., Pearce, S. F., Powell, C. Saada, A., Ohtake, A., Invernizzi, F., Lamantea, E., Sommerville, E. W., Pyle, A., Chinnery, P. F., Crushell, E., Okazaki, Y., Kohda, M., Kishita, Y., Tokuzawa, Y., Smet, J., Régal, L., Lorber, A., Khoury, A., Zeviani, M., Strom, T. M., Meitinger, T., Bertini, E. S., Van Coster, R., Klopstock, T., Haack, T. B., Minczuk, M.,** Prokisch, H.** (-) Mutations in GTPBP3 cause a mitochondrial translation defect associated with hypertrophic cardiomyopathy, lactic acidosis and encephalopathy Am J Hum Genet 95, 708-20 Vanlander, A.V., Menten, B., Smet, J., De Meirleir, L., Sante, T., De Paepe, B., Seneca, S., Pearce, S.F., Powell, C.A., Vergult, S., Michotte, A., De Latter, E., Vantomme, L., Minczuk, M. and Van Coster, R. (-) Two siblings with homozygous pathogenic splice site mutation in mitochondrial asparaginyl-tRNA synthetase (NARS2) Hum. Mutat Mutat 36, 222-231 Powell, C.A.*, Kopajtich, R.*, D'Souza, A.R., Rorbach, J., Dallabona, C., Donnini, C., Alston, C.L., Griffin, H., Pyle, A., Chinnery, P.F., Strom, T.M., Meitinger, T., Rodenburg, R.J., Schottmann, G., Schuelke, M., Romain, N., Haller, R., Ferrero, I., Haack, T.B., Taylor, R.W., Prokisch, H.**, Minczuk, M. (2015) Mutations in TRMT5 cause a defect in post-transcriptional modification of mitochondrial tRNA associated with multiple respiratory-chain deficiencies. Am J Hum Genet. 97,319-328 Coughlin, C.R. Scharer, G.H., Friederich, M.W., Yu, H.C., Geiger, E.A., Creadon-Swindell, G., Collins, A.E., Vanlander, A.V., Coster, R.V., Powell, C.A., Swanson, M.A., Minczuk, M., Van Hove, J.L., Shaikh, T.H. (2015) Mutations in the mitochondrial cysteinyl-tRNA synthase gene, CARS2, lead to a severe epileptic encephalopathy and complex movement disorder. J Med Genet. 52,532-540 and more
Start Year 2011
 
Description RNA polyadenylation in the maintenance and expression of the mitochondrial genome 
Organisation Medical Research Council (MRC)
Department MRC Functional Genomics Unit
Country United Kingdom 
Sector Public 
PI Contribution Establishing how poly(A) tails regulate mt-RNA abundance and mitochondrial protein synthesis Identifying novel proteins that play a role in mitochondrial poly(A) tail metabolism Determining whether poly(U) extensions play a role in RNA surveillance and/or turnover in human mitochondria
Collaborator Contribution Next-generation sequencing and analysis of mitochondrial RNA
Impact finantial support
Start Year 2012
 
Title GENETIC MODIFICATION OF MITOCHONDRIAL GENOMES 
Description Mutations of the mitochondrial genome (mtDNA) underlie a substantial portion of mitochondrial disease burden. These disorders are currently incurable and effectively untreatable, with heterogeneous penetrance, presentation and prognosis. To address the lack of effective treatment for these disorders, we exploited a recently developed mouse model that recapitulates common molecular features of heteroplasmic mtDNA disease in cardiac tissue: the m.5024C>T tRNAAla mouse. Through application of a programmable nuclease therapy approach, using systemically administered, mitochondrially targeted zinc-finger nucleases (mtZFN) delivered by adeno-associated virus, we induced specific elimination of mutant mtDNA across the heart, coupled to a reversion of molecular and biochemical phenotypes. These findings constitute proof of principle that mtDNA heteroplasmy correction using programmable nucleases could provide a therapeutic route for heteroplasmic mitochondrial diseases of diverse genetic origin 
IP Reference US62/646,156 
Protection Patent application published
Year Protection Granted 2018
Licensed No
Impact This technology may be useful in treating mtDNA disease.
 
Description Dissemination of scientific achievements via the internet 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Dissemination of scientific achievements and publications to the general public, scientists and others via the Unit's website, Twitter and Facebook.

https:/www.facebook.com/mrc-mbu
Twitter - @MRC_MBU
http://www.mrc-mbu.cam.ac.uk
http://www.mrc-mbu.cam.ac.uk/people/michal-minczuk
Year(s) Of Engagement Activity 2015,2016,2017,2018
URL http://www.mrc-mbu.cam.ac.uk/people/michal-minczuk
 
Description Mayfield Primary School, Cambridge 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Members of the MBU visited Mayfield Primary School, Cambridge to participate in their Science Day. This was a classroom based event, with students in Reception (aged 4 and 5 years) rotating around the room. After a short talk, the children interacted with Lego models, pipetting and colouring.
Year(s) Of Engagement Activity 2017
 
Description Opportunities Ahead 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact The event is aimed at school students in years 5 & 6 in the morning followed by students from secondary schools, sixth-forms, recent school and college leavers and parents in the afternoon. The idea is to showcase the wide variety of companies in the Cambridge area and get young people excited about their future.

Signpost 2 Skills is a new service designed to guide students from education and into working life by bringing employers into schools and students into businesses. It is funded by the Greater Cambridge Greater Peterborough LEP and the Greater Cambridge City Deal, and delivered by social enterprise, Form the Future.
Year(s) Of Engagement Activity 2017
URL http://www.mrc-mbu.cam.ac.uk/news/2768/opportunities-ahead-careers-fair-cambridge
 
Description Public Engagement Training 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact A Public Engagement Workshop entitled, "Who are the public and what do they think?" Aimed at:
Understanding body language
Intentions and measuring success
Engagement in practice - case studies/experiences
Places to find further support and methods to engage further
Year(s) Of Engagement Activity 2017
 
Description Queen Edith's Primary School, Cambridge 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Members of the MBU visited Queen Edith's Primary School, Cambridge to participate in their Science Day. This was a carousel-type event, with students in years 1-4 rotating around the room. The demonstrations included Lego models, computer games, pipetting and DNA sequencing puzzles. Feedback: Thank you so much for coming here today. The children have not stopped talking about it since leaving the room.
Year(s) Of Engagement Activity 2017
 
Description Woodhouse College Visit (to MBU) 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Eight year 12 students from Woodhouse College visited on Wednesday, 28th of June for a day focused on gene manipulation. It began with discussions surrounding the ethics and practicalities of genetic engineering followed by a practical on CRISPR-Cas9, PCR, and gel electrophoresis. This was concluded with a session to interpret the results obtained and to discuss their significance. Feedback: It was my first experience in a lab based environment and greatly broadened my perspective of the opportunities available in a career of science.It was an amazing trip, really interesting to see a research scientist's day to day life. In addition participating in a PCR and gel electrophoresis was fascinating and I received advice to help me for my university application. Thanks.The experience we got in the lab was invaluable and the techniques we used were not only relevant to our curriculum but really enjoyable. We got to explore and be a part of a really exciting, cutting-edge development in biomedical science , CRISPR- Cas9! The trip provided an incredible motivation to succeed and aim high. I am incredibly thankful for the opportunity.Visiting a working lab was an exciting opportunity and gave me a great understanding of the techniques biochemists use, including CRISPR/Cas9 which I had heard about in the media, but being so cutting edge I was surprised that the technique was so commonplace in the lab.
Year(s) Of Engagement Activity 2017,2018
 
Description Work Experience 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Visit by three Year 10 students (age 15) to the Unit for a week each - work experience/shadowing in various research groups/environments, chaperoned throughout by individual members of the Unit. Presentation by the visitors at the end of their week.
Year(s) Of Engagement Activity 2016,2017