The applicant's long-term objectives are to define the biochemical and genetic basis for human inherited neuromuscular disorders. In line with his previous work on Duchenne and Becker muscular dystrophies, the applicant proposes to apply molecular genetic, biochemical and cell biological techniques towards this goal, using diagnostic muscle biopsies, DNA, and clinical data as his primary materials. In this proposal, the applicant describes plans to continue to develop dystrophin testing as a means for biochemical diagnosis of myopathies. Dystrophin testing appears capable of detecting all Duchenne/Becker dystrophy patients even before the onset of obvious clinical symptoms, and the results are clearly prognostic. DNA testing, on the other hand, is only capable of detecting two-thirds of Duchenne/Becker patients, and the results are much less prognostic of the expected clinical severity. The proposed biochemical studies will delineate the range of clinical phenotypes possible from dystrophin abnormalities by investigating dystrophin abnormalities in limb-girdle and congenital myopathy patients. An important result of the continued patient testing will be the establishment of an extensive patient database of true limb-girdle and congenital myopathy patients. This database will be used as starting material for investigations directed towards identifying underlying biochemical defects in patients not having dystrophin abnormalities. The experimental approach to be used for identifying the protein defects in these patients will be the candidate gene/protein approach. Specifically, we have recently developed experimental methods by which we can clone human genes and produce large quantities of the corresponding human protein from any published vertebrate sequence with only a few hours of work. Antibodies raised against the produced proteins have proven to highly sensitive and specifically directed against the human protein. In this proposal we describe our application of these techniques towards two human proteins, the chromosome-6 dystrophin-related protein, and the skeletal muscle sodium channel. The cloned human genes and the corresponding antibodies will be used to investigate the possible involvement of these proteins in neuromuscular disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
1R01NS029525-01
Application #
3416375
Study Section
Molecular Cytology Study Section (CTY)
Project Start
1991-01-01
Project End
1994-12-31
Budget Start
1991-01-01
Budget End
1991-12-31
Support Year
1
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Pittsburgh
Department
Type
Schools of Medicine
DUNS #
053785812
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Punetha, Jaya; Kesari, Akanchha; Hoffman, Eric P et al. (2017) Novel Col12A1 variant expands the clinical picture of congenital myopathies with extracellular matrix defects. Muscle Nerve 55:277-281
Perovanovic, Jelena; Dell'Orso, Stefania; Gnochi, Viola F et al. (2016) Laminopathies disrupt epigenomic developmental programs and cell fate. Sci Transl Med 8:335ra58
Chen, Xi; Jung, Jin-Gyoung; Shajahan-Haq, Ayesha N et al. (2016) ChIP-BIT: Bayesian inference of target genes using a novel joint probabilistic model of ChIP-seq profiles. Nucleic Acids Res 44:e65
Punetha, Jaya; Mansoor, Simin; Bertorini, Tulio E et al. (2016) Somatic mosaicism due to a reversion variant causing hemi-atrophy: a novel variant of dystrophinopathy. Eur J Hum Genet 24:1511-4
Willkomm, Lena; Heredia, Raul; Hoffmann, Katrin et al. (2016) Homozygous mutation in Atlastin GTPase 1 causes recessive hereditary spastic paraplegia. J Hum Genet 61:571-3
O'Grady, Gina L; Lek, Monkol; Lamande, Shireen R et al. (2016) Diagnosis and etiology of congenital muscular dystrophy: We are halfway there. Ann Neurol 80:101-11
Punetha, Jaya; Kesari, Akanchha; Uapinyoying, Prech et al. (2016) Targeted Re-Sequencing Emulsion PCR Panel for Myopathies: Results in 94 Cases. J Neuromuscul Dis 3:209-225
Wang, Niya; Hoffman, Eric P; Chen, Lulu et al. (2016) Mathematical modelling of transcriptional heterogeneity identifies novel markers and subpopulations in complex tissues. Sci Rep 6:18909
Many, Gina M; Yokosaki, Yasuyuki; Uaesoontrachoon, Kitipong et al. (2016) OPN-a induces muscle inflammation by increasing recruitment and activation of pro-inflammatory macrophages. Exp Physiol 101:1285-1300
Wang, Niya; Gong, Ting; Clarke, Robert et al. (2015) UNDO: a Bioconductor R package for unsupervised deconvolution of mixed gene expressions in tumor samples. Bioinformatics 31:137-9

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