This application addresses Notice Number (NOT-OD-09-058) and Notice Title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications, and asks for competitive revision of R01DK78755. The acyl-CoA dehydrogenases (ACDs) are a family of multimeric flavoenzymes that catalyze the 1,2-dehydrogenation of acyl-CoA esters in fatty acid 2-oxidation and amino acid catabolism. Inborn errors of metabolism have been identified in seven of the ACDs. The long range objective of this project has been to investigate important structure/function relationships in the ACD gene family. We have described and characterized several new members of the ACD gene family. Among these are 3 enzymes with significant activities with long chain substrates: long and very long chain acyl-CoA dehydrogenases (LCAD and VLCAD, respectively), and ACD9 and. Our prior and preliminary studies show that these enzymes have distinct substrate utilization profiles, tissue and developmental expression patterns, exist in multiple active forms in the cell, and are present in multiple subcellular locations. The goal of the funded application is to characterize the physiologic roles of LCAD, VLCAD, and ACD9 and explore the ramifications of genetic deficiencies of these enzymes in humans and mouse models.
Specific Aim 2 of the original application is to more completely characterize ACD9 and its deficiency in humans.
Specific Aim 2 a is identification of additional patients with ACD9 deficiency and definition of its clinical spectrum.
Specific Aim 2 b is characterization of the subcellular distribution of ACD9 and the function and molecular configuration of ACD9 protein outside of mitochondria. I hypothesize that this alternative form of ACD9 has non-enzymatic """"""""moonlighting"""""""" functions in the cell. I am requesting a supplement to the application to extend my ability to address these aims. First, I have identified more potential deficient patients than originally anticipated. I am requesting funds to support additional technician time to process these samples more quickly and accelerate our progress. Secondly, our understanding of the pathophysiology of ACAD9 deficiency and our ability to study the moonlighting functions of this gene product are limited by the availability of only tissue culture cells with ACAD9 deficiency. To augment these studies, I am requesting supplemental funds to purchase an ACAD9 knock out mouse from the Texas A&M Institute for Genome Medicine, which has it available in its catalogue. The biochemical and pathophysiologic phenotype of the animal will be characterized including metabolic profiling, complete histologic survey, and physiologic effects of the deficiency on the animal under basal and resting conditions. Immunohistochemistry and subcellular fractionation studies will examine for presences of ACAD9 in non-mitochondrial locations and loss in the knock out mouse model.

Public Health Relevance

The acyl-CoA dehydrogenases are important enzymes in maintaining normal chemical balance in the body. We have identified a new genetic disorder of one of these enzymes that leads to liver failure. Studying this disorder is important to learn more about its clinical presentation and treatment

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
3R01DK078775-02S1
Application #
7810760
Study Section
Special Emphasis Panel (ZRG1-GGG-M (95))
Program Officer
Mckeon, Catherine T
Project Start
2009-09-20
Project End
2011-08-31
Budget Start
2009-09-20
Budget End
2011-08-31
Support Year
2
Fiscal Year
2009
Total Cost
$439,048
Indirect Cost
Name
University of Pittsburgh
Department
Pediatrics
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Leipnitz, Guilhian; Mohsen, Al-Walid; Karunanidhi, Anuradha et al. (2018) Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency. Sci Rep 8:1165
Repp, Birgit M; Mastantuono, Elisa; Alston, Charlotte L et al. (2018) Clinical, biochemical and genetic spectrum of 70 patients with ACAD9 deficiency: is riboflavin supplementation effective? Orphanet J Rare Dis 13:120
Gillingham, Melanie B; Heitner, Stephen B; Martin, Julie et al. (2017) Triheptanoin versus trioctanoin for long-chain fatty acid oxidation disorders: a double blinded, randomized controlled trial. J Inherit Metab Dis 40:831-843
Pena, Loren D M; van Calcar, Sandra C; Hansen, Joyanna et al. (2016) Outcomes and genotype-phenotype correlations in 52 individuals with VLCAD deficiency diagnosed by NBS and enrolled in the IBEM-IS database. Mol Genet Metab 118:272-81
Vockley, J; Charrow, J; Ganesh, J et al. (2016) Triheptanoin treatment in patients with pediatric cardiomyopathy associated with long chain-fatty acid oxidation disorders. Mol Genet Metab 119:223-231
Staufner, Christian; Haack, Tobias B; Köpke, Marlies G et al. (2016) Recurrent acute liver failure due to NBAS deficiency: phenotypic spectrum, disease mechanisms, and therapeutic concepts. J Inherit Metab Dis 39:3-16
Prabhu, Dolly; Goldstein, Amy C; El-Khoury, Riyad et al. (2015) ANT2-defective fibroblasts exhibit normal mitochondrial bioenergetics. Mol Genet Metab Rep 3:43-46
Vockley, Jerry; Marsden, Deborah; McCracken, Elizabeth et al. (2015) Long-term major clinical outcomes in patients with long chain fatty acid oxidation disorders before and after transition to triheptanoin treatment--A retrospective chart review. Mol Genet Metab 116:53-60
Schiff, Manuel; Haberberger, Birgit; Xia, Chuanwu et al. (2015) Complex I assembly function and fatty acid oxidation enzyme activity of ACAD9 both contribute to disease severity in ACAD9 deficiency. Hum Mol Genet 24:3238-47
Edmunds, Lia R; Sharma, Lokendra; Kang, Audry et al. (2014) c-Myc programs fatty acid metabolism and dictates acetyl-CoA abundance and fate. J Biol Chem 289:25382-92

Showing the most recent 10 out of 31 publications