TY - JOUR
T1 - Loss of LMOD1 impairs smooth muscle cytocontractility and causes megacystis microcolon intestinal hypoperistalsis syndrome in humans and mice
AU - Halim, Danny
AU - Wilson, Michael P.
AU - Oliver, Daniel
AU - Brosens, Erwin
AU - Verheij, Joke B.G.M.
AU - Han, Yu
AU - Nanda, Vivek
AU - Lyu, Qing
AU - Doukas, Michael
AU - Stoop, Hans
AU - Brouwer, Rutger W.W.
AU - Van Ijcken, Wilfred F.J.
AU - Slivano, Orazio J.
AU - Burns, Alan J.
AU - Christie, Christine K.
AU - De Mesy Bentley, Karen L.
AU - Brooks, Alice S.
AU - Tibboel, Dick
AU - Xu, Suowen
AU - Jin, Zheng Gen
AU - Djuwantono, Tono
AU - Yan, Wei
AU - Alves, Maria M.
AU - Hofstra, Robert M.W.
AU - Miano, Joseph M.
N1 - Funding Information:
The two-component CRISPR knockout of Lmod1 was conducted in the Genome Editing Core of the University of Nevada, Reno School of Medicine, which is supported, in part, by a COBRE grant from the NIH (1P30GM110767). J.M.M. is supported by NIH Grants HL-117907 and HL-112793.
PY - 2017/3/28
Y1 - 2017/3/28
N2 - Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is a congenital visceral myopathy characterized by severe dilation of the urinary bladder and defective intestinal motility. The genetic basis of MMIHS has been ascribed to spontaneous and autosomal dominant mutations in actin gamma 2 (ACTG2), a smooth muscle contractile gene. However, evidence suggesting a recessive origin of the disease also exists. Using combined homozygosity mapping and whole exome sequencing, a genetically isolated family was found to carry a premature termination codon in Leiomodin1 (LMOD1), a gene preferentially expressed in vascular and visceral smooth muscle cells. Parents heterozygous for the mutation exhibited no abnormalities, but a child homozygous for the premature termination codon displayed symptoms consistent with MMIHS. We used CRISPR-Cas9 (CRISPR-associated protein) genome editing of Lmod1 to generate a similar premature termination codon. Mice homozygous for the mutation showed loss of LMOD1 protein and pathology consistent with MMIHS, including late gestation expansion of the bladder, hydronephrosis, and rapid demise after parturition. Loss of LMOD1 resulted in a reduction of filamentous actin, elongated cytoskeletal dense bodies, and impaired intestinal smooth muscle contractility. These results define LMOD1 as a disease gene for MMIHS and suggest its role in establishing normal smooth muscle cytoskeletal and contractile coupling.
AB - Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is a congenital visceral myopathy characterized by severe dilation of the urinary bladder and defective intestinal motility. The genetic basis of MMIHS has been ascribed to spontaneous and autosomal dominant mutations in actin gamma 2 (ACTG2), a smooth muscle contractile gene. However, evidence suggesting a recessive origin of the disease also exists. Using combined homozygosity mapping and whole exome sequencing, a genetically isolated family was found to carry a premature termination codon in Leiomodin1 (LMOD1), a gene preferentially expressed in vascular and visceral smooth muscle cells. Parents heterozygous for the mutation exhibited no abnormalities, but a child homozygous for the premature termination codon displayed symptoms consistent with MMIHS. We used CRISPR-Cas9 (CRISPR-associated protein) genome editing of Lmod1 to generate a similar premature termination codon. Mice homozygous for the mutation showed loss of LMOD1 protein and pathology consistent with MMIHS, including late gestation expansion of the bladder, hydronephrosis, and rapid demise after parturition. Loss of LMOD1 resulted in a reduction of filamentous actin, elongated cytoskeletal dense bodies, and impaired intestinal smooth muscle contractility. These results define LMOD1 as a disease gene for MMIHS and suggest its role in establishing normal smooth muscle cytoskeletal and contractile coupling.
KW - CRISPR-Cas9
KW - Genetics
KW - Leiomodin
KW - Myopathy
KW - Smooth muscle
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U2 - 10.1073/pnas.1620507114
DO - 10.1073/pnas.1620507114
M3 - Article
C2 - 28292896
AN - SCOPUS:85016429718
SN - 0027-8424
VL - 114
SP - E2739-E2747
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 13
ER -