Natural Product Screening Reveals Naphthoquinone Complex I Bypass Factors.
Deficiency of mitochondrial complex I is encountered in both rare and common diseases, but we have limited therapeutic options to treat this lesion to the oxidative phosphorylation system (OXPHOS). Idebenone and menadione are redox-active molecules capable of rescuing OXPHOS activity by engaging complex I-independent pathways of entry, often referred to as "complex I bypass." In the present study, we created a cellular model of complex I deficiency by using CRISPR genome editing to knock out Ndufa9 in mouse myoblasts, and utilized this cell line to develop a high-throughput screening platform for novel complex I bypass factors. We screened a library of ~40,000 natural product extracts and performed bioassay-guided fractionation on a subset of the top scoring hits. We isolated four plant-derived 1,4-naphthoquinone complex I bypass factors with structural similarity to menadione: chimaphilin and 3-chloro-chimaphilin from Chimaphila umbellata and dehydro-α-lapachone and dehydroiso-α-lapachone from Stereospermum euphoroides. We also tested a small number of structurally related naphthoquinones from commercial sources and identified two additional compounds with complex I bypass activity: 2-methoxy-1,4-naphthoquinone and 2-methoxy-3-methyl-1,4,-naphthoquinone. The six novel complex I bypass factors reported here expand this class of molecules and will be useful as tool compounds for investigating complex I disease biology.
Howard Hughes Medical Institute and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA, United States of America.|Broad Institute of MIT and Harvard, Cambridge, MA, United States of America.
Scott B
http://orcid.org/0000-0003-1038-9614
SB
Vafai
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, United States of America.
Emily
E
Mevers
Howard Hughes Medical Institute and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA, United States of America.|Broad Institute of MIT and Harvard, Cambridge, MA, United States of America.
Kathleen W
KW
Higgins
Howard Hughes Medical Institute and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA, United States of America.|Broad Institute of MIT and Harvard, Cambridge, MA, United States of America.
Yevgenia
Y
Fomina
Department of Dermatology, Massachusetts General Hospital, Boston, MA, United States of America.
Jianming
J
Zhang
Department of Dermatology, Massachusetts General Hospital, Boston, MA, United States of America.
Anna
A
Mandinova
Natural Products Branch, National Cancer Institute, Frederick, MD, United States of America.
David
D
Newman
Center for Systems Biology, Massachusetts General Hospital, Boston, MA, United States of America.
Stanley Y
SY
Shaw
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, United States of America.
Jon
J
Clardy
Howard Hughes Medical Institute and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA, United States of America.|Broad Institute of MIT and Harvard, Cambridge, MA, United States of America.|Department of Systems Biology, Harvard Medical School, Boston, MA, United States of America.
Vamsi K
VK
Mootha
D000818:Animals; D029402:Bignoniaceae / Q000737:chemistry; D001688:Biological Products / Q000737:chemistry / Q000494:pharmacology*; D064113:CRISPR-Cas Systems; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D042967:Electron Transport Complex I / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D029791:Ericaceae / Q000737:chemistry; D000072669:Gene Editing; D055786:Gene Knockout Techniques; D057166:High-Throughput Screening Assays; D051379:Mice; D032446:Myoblasts / Q000187:drug effects / Q000378:metabolism; D009285:Naphthoquinones / Q000737:chemistry / Q000494:pharmacology*; D010085:Oxidative Phosphorylation; D023701:Smegmamorpha / Q000378:metabolism
D001688:Biological Products; D009285:Naphthoquinones; D042967:Electron Transport Complex I