Mitochondrial Respiratory Measurements in Patient-derived Fibroblasts

Prashant Mishra; Ting Zhang; Ming Guo; David Chan

doi:10.21769/BioProtoc.3446

Mitochondrial Respiratory Measurements in Patient-derived Fibroblasts

Bio Protoc. 2019 Dec 5;9(23):e3446. doi: 10.21769/BioProtoc.3446.

Authors

Prashant Mishra¹, Ting Zhang², Ming Guo^{2

3

4}, David Chan¹

Affiliations

¹ Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
² Department of Neurology, UCLA David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
³ Department of Molecular and Medical Pharmacology, UCLA David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
⁴ California Nanosystems Institute at UCLA, Los Angeles, CA 90095, USA.

Abstract

Mitochondrial dysfunction is associated with a number of human diseases. As an example, we recently established in vivo Drosophila models of IBMPFD (Inclusion body myopathy, Paget disease, and frontotemporal dementia), and uncovered that human disease mutations of the p97/VCP (Valosin Containing Protein) gene behave as hyperactive alleles associated with mitochondrial defects. Pharmacologic inhibition of VCP strongly suppressed disease and mitochondrial pathology in these animal models. In this protocol, we describe a method to evaluate mitochondrial respiratory function in IBMPFD patient-derived fibroblasts, as well as investigate the role of pharmacologic treatments. These experiments complement work done in animal models by investigating mitochondrial biology and the pharmacologic response in a human cell-based model of the disease. In principle, this technique can be used to investigate mitochondrial respiratory function for any disease in which patient-derived fibroblasts are available.

Keywords: Inclusion body myopathy; Mitochondrial respiration; Paget disease and frontotemporal dementia (IBMPFD); Patient-derived fibroblasts; Seahorse XF assay; VCP/p97.