1. Anatomical Laser Microdissection of the Ileum Reveals mtDNA Depletion Recovery in A Mitochondrial Neuro-Gastrointestinal Encephalomyopathy (MNGIE) Patient Receiving Liver Transplant
- Author
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Elisa Boschetti, Leonardo Caporali, Roberto D’Angelo, Carolina Malagelada, Anna Accarino, Maria Teresa Dotti, Roberta Costa, Giovanna Cenacchi, Loris Pironi, Rita Rinaldi, Vincenzo Stanghellini, Stefano Ratti, Lucia Manzoli, Valerio Carelli, Roberto De Giorgio, Institut Català de la Salut, [Boschetti E] Department of Biomedical and Neuromotor Sciences (DIBINEM), University of Bologna, Bologna, Italy. IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy. [Caporali L] IRCCS Istituto delle Scienze Neurologiche di Bologna, Bologna, Italy. [D'Angelo R] IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy. [Malagelada C, Accarino A] Centro de Investigacion Biomedica en Red de Enfermedades Hepáticas y Digestivas (CIBEREHD), Barcelona, Spain. Vall d’Hebron Hospital Universitari, Barcelona, Spain. Departament de Medicina, Universitat Autònoma de Barcelona, Bellaterra, Spain. [Dotti MT] Department of Medical, Surgical and Neurological Sciences, University of Siena, Siena, Italy, Vall d'Hebron Barcelona Hospital Campus, Boschetti, Elisa, Caporali, Leonardo, D'Angelo, Roberto, Malagelada, Carolina, Accarino, Anna, Dotti, Maria Teresa, Costa, Roberta, Cenacchi, Giovanna, Pironi, Lori, Rinaldi, Rita, Stanghellini, Vincenzo, Ratti, Stefano, Manzoli, Lucia, Carelli, Valerio, and De Giorgio, Roberto
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gastrointestinal degeneration ,Investigative Techniques::Investigative Techniques::Micromanipulation::Microdissection::Laser Capture Microdissection [ANALYTICAL, DIAGNOSTIC AND THERAPEUTIC TECHNIQUES, AND EQUIPMENT] ,Laser ,Fetge - Trasplantació ,ADN mitocondrial ,Laser Capture Microdissection ,Encefalomielitis ,DNA, Mitochondrial ,Catalysis ,Inorganic Chemistry ,Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::DNA::DNA, Circular::DNA, Mitochondrial [CHEMICALS AND DRUGS] ,Ileum ,Mitochondrial Encephalomyopathies ,nucleótidos y nucleósidos de ácidos nucleicos::ácidos nucleicos::ADN::ADN circular::ADN mitocondrial [COMPUESTOS QUÍMICOS Y DROGAS] ,mitochondrial disorder ,Humans ,Physical and Theoretical Chemistry ,Molecular Biology ,Spectroscopy ,enfermedades del sistema nervioso::enfermedades del sistema nervioso central::enfermedades cerebrales::enfermedades cerebrales metabólicas::enfermedades del sistema nervioso::encefalomiopatías mitocondriales [ENFERMEDADES] ,terapéutica::terapia biológica::tratamientos basados en células y tejidos::trasplante de tejidos::trasplante de hígado [TÉCNICAS Y EQUIPOS ANALÍTICOS, DIAGNÓSTICOS Y TERAPÉUTICOS] ,microanatomical dissection ,mtDNA depletion ,Nervous System Diseases::Central Nervous System Diseases::Brain Diseases::Brain Diseases, Metabolic::Nervous System Diseases::Mitochondrial Encephalomyopathies [DISEASES] ,Lasers ,Organic Chemistry ,General Medicine ,mitochondrial disorders ,MNGIE ,Liver Transplantation ,Computer Science Applications ,técnicas de investigación::técnicas de investigación::micromanipulación::microdisección::microdisección por láser [TÉCNICAS Y EQUIPOS ANALÍTICOS, DIAGNÓSTICOS Y TERAPÉUTICOS] ,Therapeutics::Biological Therapy::Cell- and Tissue-Based Therapy::Tissue Transplantation::Liver Transplantation [ANALYTICAL, DIAGNOSTIC AND THERAPEUTIC TECHNIQUES, AND EQUIPMENT] ,Mitocondris - Malalties ,Metabolism, Inborn Error ,Metabolism, Inborn Errors ,Human - Abstract
Microanatomical dissection; Mitochondrial disorders; MtDNA depletion Disección microanatómica; Trastornos mitocondriales; Agotamiento del ADNmt Dissecció microanatòmica; Trastorns mitocondrials; Esgotament de l'ADNmt Mitochondrial neuro-gastrointestinal encephalomyopathy (MNGIE) is a rare genetic disorder characterized by thymidine phosphorylase (TP) enzyme defect. The absence of TP activity induces the imbalance of mitochondrial nucleotide pool, leading to impaired mitochondrial DNA (mtDNA) replication and depletion. Since mtDNA is required to ensure oxidative phosphorylation, metabolically active tissues may not achieve sufficient energy production. The only effective life-saving approach in MNGIE has been the permanent replacement of TP via allogeneic hematopoietic stem cell or liver transplantation. However, the follow-up of transplanted patients showed that gut tissue changes do not revert and fatal complications, such as massive gastrointestinal bleeding, can occur. The purpose of this study was to clarify whether the reintroduction of TP after transplant can recover mtDNA copy number in a normal range. Using laser capture microdissection and droplet-digital-PCR, we assessed the mtDNA copy number in each layer of full-thickness ileal samples of a naive MNGIE cohort vs. controls and in a patient pre- and post-TP replacement. The treatment led to a significant recovery of gut tissue mtDNA amount, thus showing its efficacy. Our results indicate that a timely TP replacement is needed to maximize therapeutic success before irreversible degenerative tissue changes occur in MNGIE. The work was supported by Ministero dell’Istruzione, dell’Università e della Ricerca-Dipartimenti eccellenti on the Project Personalized medicine. LC and VC are supported by the Italian Ministry of Health (Ricerca Corrente 2021 funding). RDG is supported by funds from the University of Ferrara.
- Published
- 2022
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