Cammarato Lab
The Cammarato Lab is located in the Division of Cardiology in the Department of Medicine at The Johns Hopkins University School of Medicine. We are interested in basic mechanisms of striated muscle biology. We employ an array of imaging techniques to study “structural physiology” of cardiac and skeletal muscle. Drosophila melanogaster, the fruit fly, expresses both forms of striated muscle and benefits greatly from powerful genetic tools. We investigate conserved myopathic (muscle disease) processes and perform hierarchical and integrative analysis of muscle function from the level of single molecules and macromolecular complexes through the level of the tissue itself.
Projects
Active projects within the Cammarato Lab are funded by the NIH/NHLBI, NIH/NIA, and NIH/NIGMS.
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1R01HL124091 (Cammarato, P.I.)
4/01/15-3/31/20
NIH/NHLBI
Pathogenesis and in vivo suppression of thin filament based cardiomyopathiesHere, we propose to use a transgenic animal model system, Drosophila melanogaster (the fruit fly), to define the mechanisms by which mutations in various thin filament components lead to human cardiac disease. We will produce several new models of actin and troponin-T-based cardiomyopathies to determine the molecular defects that drive diverse and complex tissue remodeling. Finally, in vivo genetic suppression experiments, designed to ameliorate cardiac decline during troponin-T-mediated disease, will resolve novel interactions among thin filament components involved in regulating muscle contraction.
R01 GM32443 (Bernstein, P.I.)
5/01/14-4/30/16
SDSU/NIH/NIGMS
Genetics and Molecular Biology of Striated Muscle MyosinThe goal of this project is to examine the mechanism by which the myosin molecular motor functions in striated muscle. Using a transgenic model system, the fruit fly Drosophila melanogaster, we will produce models of human myosin-based muscle disease (distal arthrogryposis) and heart disease (hypertrophic cardiomyopathy) to determine the molecular defects that cause abnormal skeletal and cardiac muscle function. We will use genetic suppression and drug treatment to better understand the bases of myosin malfunction and to develop our model system as a means of testing potential therapeutic approaches.
Role: Collaborator
R01 AG045428 (Engler, P.I.)
8/01/13-6/30/18
UCSD/NIH/NIA
Mechanogenetics: An Integrated Approach to Aging in Muscle DysfunctionThe goal of this project is to examine how specific intercalated disc proteins, which are upregulated with age as a part of a genotype, alter the age-associated performance and mechanical stiffness of the Drosophila heart tube. Data from this rapidly aging, genetically tractable model system will be correlated with aged mammalian systems (with and without dysfunction) to assess the predictive power of the fly model in determining age-associated negative changes in heart performance.
Role: Collaborator
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AFAR Research Grant
Cammarato, A., P.I.
07/01/12-06/30/14
American Federation for Aging ResearchDetermining and manipulating age-dependent changes in myocardial stiffness, in vivo. The specific aims of the project are: 1) Identify differences in potential collective, age-related myocardial stiffening events among multiple Drosophila control lines. We will ascertain passive mechanical differences in young vs. old fly hearts from several control strains including Oregon-R, w1118, Canton-S and GMH5-GAL4 x yw. 2) Investigate the effect of aging on passive mechanical properties of Drosophila myocardium with cardiac-specific overexpression of the transcription factor FOXO, which is known to rejuvenate cardiac performance and promote muscle proteostasis in senescent flies. 3) Employ targeted, cardiac-specific RNA interference to improve morphology, performance and mechanics of senescent hearts.
Scientist Development Grant
Cammarato, A., P.I.
07/01/10-06/30/14
American Heart Association (AHA) National Center, 10SDG4180089
Identifying and manipulating age- and mutation-dependent modifiers of cardiac function using the Drosophila model. The specific aims of the project are: 1) Identify age-related changes in cardiac expression profiles using GeneChip microarrays that represent the complete Drosophila genome. This will be done for young and old wildtype fly hearts and for the hearts of two myosin heavy chain mutants and a troponin mutant. 2) Employ RNA interference to knockdown specific cardiac transcripts to determine their role in myopathic responses to the myosin mutations.
Lab Members
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Assistant Professor of Cardiology
Education and Training
- 1995 — Bachelor of Science, Biology
Mary Washington College, Fredericksburg, VA - 2004 — Doctor of Philosophy, Physiology and Biophysics
Boston University School of Medicine, Boston, MA - 2004 - 2008 — Postdoctoral Fellow, Molecular Genetics, Structural Biology, Cardiac Function, San Diego State University, San Diego, CA
Positions and Employment
- 1995 — General Biology Lab Instructor, Dept of Biology, University of Delaware, Newark, DE
- 2004 - 2011 — Faculty Adjunct, Dept of Biology, San Diego State University, San Diego, CA
- 2006 - 2008 — American Heart Association Post-Doctoral Fellow, Dept of Biology, San Diego State University, San Diego, CA
- 2009 - 2010 — Staff Scientist, Sanford-Burnham Medical Research Institute, NASCR, La Jolla, CA
- 2010 - 2011 — Research Assistant Professor, Sanford-Burnham Medical Research Institute, NASCR, La Jolla, CA
- 2011 - Present — Assistant Professor, Johns Hopkins University School of Medicine, Department of Medicine, Division of Cardiology, Baltimore, MD
Professional Memberships
- 1999 - present — The American Physiological Society
- 2000 - present — Biophysical Society
- 2006 - present — The American Society for Cell Biology
- 2012 - present — Genetics Society of America
- 1995 — Bachelor of Science, Biology
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Research Specialist, Lab Manager
Education and Training
- 2003 - 2007 — B.S Physician Assistant. Birla Institute of Technology and Science
- 2009 - 2012 — M.S in Biology (with emphasis in cell and molecular biology), San Diego State University, CA, USA. Thesis: Studying human myosin storage myopathy using a Drosophila model
Positions and Employment
- 2009 - 2012 — Graduate Research Assistant, San Diego State University, San Diego, CA
- 2009 - 2012 — Graduate Teaching Assistant, San Diego State University, San Diego, CA
- 2012 - Present — Research Specialist, Lab Manager, Johns Hopkins University School of Medicine, Department of Medicine, Division of Cardiology, Baltimore, MD
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Postdoctoral Fellow
Education and Training
- 2007 — Bachelor of Science, Chemical Biology, Stevens Institute of Technology, Hoboken, NJ
- 2014 — Doctor of Philosophy, Physiology, Boston University, Boston, MA
- 2014 - Present — Postdoctoral Fellow, Molecular mechanisms of cardiac contraction, Johns Hopkins University, Baltimore, MD
Positions and Employment
- 2007 - 2014 — Ph.D. student, Boston University, Boston, MA
- 2014 - Present — Postdoctoral Fellow, Johns Hopkins University School of Medicine, Department of Medicine, Division of Cardiology, Baltimore, MD
Professional Memberships
- 2007 - 2015 — Biophysical Society
- 2015 - present — American Heart Association
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Postdoctoral Fellow
Education and Training
- 2007 — Bachelor of Science, Biochemistry, Delhi University, India
- 2009 — Master of Science, Biological Sciences, Indian Institute of Sciences, Bangalore, India
- 2016 — Doctor of Philosophy, Muscle Genetics, Indian Institute of Science, Bangalore, India
- 2016 - present — Postdoctoral Fellow, Johns Hopkins University School of Medicine, Department of Medicine, Division of Cardiology, Baltimore, MD
Positions and Employment
- 2009 - 2016 — Ph.D. student, Indian Institute of Science, Bangalore, India
- 2016 - present — Postdoctoral Fellow, Johns Hopkins University School of Medicine, Department of Medicine, Division of Cardiology, Baltimore, MD
Professional Memberships
- 2008 - present — Society of Biological Chemists, India
Collaborators
- Ivor J. Benjamin, MD, FAHA, FACC
Cardiovascular Center
Medical College of Wisconsin - Sanford I. Bernstein, Ph.D.
Department of Biology
San Diego State University - Adam J. Engler, Ph.D.
Department of Bioengineering
University of California, San Diego - D. Brian Foster, Ph.D.
Department of Medicine, Division of Cardiology
Johns Hopkins University School of Medicine
- David A. Kass, M.D.
Department of Medicine, Division of Cardiology
Johns Hopkins University School of Medicine - William J. Lehman, Ph.D.
Department of Physiology and Biophysics
Boston University School of Medicine - Karen Ocorr Ph.D. and Rolf Bodmer, Ph.D.
Development and Aging Program
Sanford Burnham Medical Research Institute - John Sparrow, D. Phil.
Department of Biology
University of York
- Douglas M. Swank Ph.D.
Biology Department
Rensselaer Polytechnic Institute - J. Paul Taylor, M.D., Ph.D.
Developmental Neurobiology
St. Jude Children's Research Hospital - Larry Tobacman, M.D.
Cardiology
University of Illinois College of Medicine - Jennifer Van Eyk, Ph.D.
Cardiology, Biol. Chem. and Biomed. Eng
Johns Hopkins University
Publications
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1: Cammarato A, Hatch V, Saide J, Craig R, Sparrow JC, Tobacman LS, Lehman W. Drosophila muscle regulation characterized by electron microscopy and three-dimensional reconstruction of thin filament mutants. Biophys J. 2004 Mar;86(3):1618-24. PubMed PMID: 14990488; PubMed Central PMCID: PMC1303996.
2: Ayme-Southgate A, Saide J, Southgate R, Bounaix C, Cammarato A, Patel S, Wussler C. In indirect flight muscles Drosophila projectin has a short PEVK domain, and its NH2-terminus is embedded at the Z-band. J Muscle Res Cell Motil. 2005;26(6-8):467-77. PubMed PMID: 16465474.
3: Cammarato A, Craig R, Sparrow JC, Lehman W. E93K charge reversal on actin perturbs steric regulation of thin filaments. J Mol Biol. 2005 Apr 15;347(5):889-94. PubMed PMID: 15784249.
4: Melkani GC, Cammarato A, Bernstein SI. alphaB-crystallin maintains skeletal muscle myosin enzymatic activity and prevents its aggregation under heat-shock stress. J Mol Biol. 2006 May 5;358(3):635-45. Epub 2006 Mar 3. PubMed PMID: 16546210.
5: Suggs JA, Cammarato A, Kronert WA, Nikkhoy M, Dambacher CM, Megighian A, Bernstein SI. Alternative S2 hinge regions of the myosin rod differentially affect muscle function, myofibril dimensions and myosin tail length. J Mol Biol. 2007 Apr 13;367(5):1312-29. Epub 2007 Jan 23. PubMed PMID: 17316684; PubMed Central PMCID: PMC1965590.
6: Cammarato A, Dambacher CM, Knowles AF, Kronert WA, Bodmer R, Ocorr K, Bernstein SI. Myosin transducer mutations differentially affect motor function, myofibril structure, and the performance of skeletal and cardiac muscles. Mol Biol Cell. 2008 Feb;19(2):553-62. Epub 2007 Nov 28. PubMed PMID: 18045988; PubMed Central PMCID: PMC2230588.
7: Ocorr K, Fink M, Cammarato A, Bernstein S, Bodmer R. Semi-automated Optical Heartbeat Analysis of small hearts. J Vis Exp. 2009 Sep 16;(31). pii: 1435. doi: 10.3791/1435. PubMed PMID: 19759521; PubMed Central PMCID: PMC3150057.
8: Alayari NN, Vogler G, Taghli-Lamallem O, Ocorr K, Bodmer R, Cammarato A*. Fluorescent labeling of Drosophila heart structures. J Vis Exp. 2009 Oct 13;(32). pii: 1423. doi: 10.3791/1423. PubMed PMID: 19826399; PubMed Central PMCID: PMC3164059. (* corresponding author)
9: Cammarato A*, Craig R, Lehman W. Electron microscopy and three-dimensional reconstruction of native thin filaments reveal species-specific differences in regulatory strand densities. Biochem Biophys Res Commun. 2010 Jan 1;391(1):193-7. doi: 10.1016/j.bbrc.2009.11.030. Epub 2009 Nov 10. PubMed PMID: 19900412; PubMed Central PMCID: PMC2818542. (* corresponding author)
10: Neely GG*, Kuba K*, Cammarato A*, Isobe K, Amann S, Zhang L, Murata M, Elmén L, Gupta V, Arora S, Sarangi R, Dan D, Fujisawa S, Usami T, Xia CP, Keene AC, Alayari NN, Yamakawa H, Elling U, Berger C, Novatchkova M, Koglgruber R, Fukuda K, Nishina H, Isobe M, Pospisilik JA, Imai Y, Pfeufer A, Hicks AA, Pramstaller PP, Subramaniam S, Kimura A, Ocorr K, Bodmer R, Penninger JM. A global in vivo Drosophila RNAi screen identifies NOT3 as a conserved regulator of heart function. Cell. 2010 Apr 2;141(1):142-53. doi: 10.1016/j.cell.2010.02.023. PubMed PMID: 20371351; PubMed Central PMCID: PMC2855221. (* equally contributing authors)
11: Melkani GC, Lee CF, Cammarato A, Bernstein SI. Drosophila UNC-45 prevents heat-induced aggregation of skeletal muscle myosin and facilitates refolding of citrate synthase. Biochem Biophys Res Commun. 2010 May 28;396(2):317-22. doi: 10.1016/j.bbrc.2010.04.090. Epub 2010 Apr 18. PubMed PMID: 20403336; PubMed Central PMCID: PMC2888609.
12: Sousa D, Cammarato A, Jang K, Graceffa P, Tobacman LS, Li XE, Lehman W. Electron microscopy and persistence length analysis of semi-rigid smooth muscle tropomyosin strands. Biophys J. 2010 Aug 4;99(3):862-8. doi: 10.1016/j.bpj.2010.05.004. PubMed PMID: 20682264; PubMed Central PMCID: PMC2913205.
13: Vikhorev PG, Vikhoreva NN, Cammarato A, Sparrow JC. In vitro motility of native thin filaments from Drosophila indirect flight muscles reveals that the held-up 2 TnI mutation affects calcium activation. J Muscle Res Cell Motil. 2010 Sep;31(3):171-9. doi: 10.1007/s10974-010-9221-x. Epub 2010 Jul 24. PubMed PMID: 20658179.
14: Cammarato A, Ahrens CH, Alayari NN, Qeli E, Rucker J, Reedy MC, Zmasek CM, Gucek M, Cole RN, Van Eyk JE, Bodmer R, O'Rourke B, Bernstein SI, Foster DB. A mighty small heart: the cardiac proteome of adult Drosophila melanogaster. PLoS One. 2011 Apr 25;6(4):e18497. doi: 10.1371/journal.pone.0018497. PubMed PMID: 21541028; PubMed Central PMCID: PMC3081823.
15: Kaushik G, Fuhrmann A, Cammarato A*, Engler AJ*. In situ mechanical analysis of myofibrillar perturbation and aging on soft, bilayered Drosophila myocardium. Biophys J. 2011 Dec 7;101(11):2629-37. doi: 10.1016/j.bpj.2011.10.042. PubMed PMID: 22261050; PubMed Central PMCID: PMC3297788. (* co-corresponding authors)
16: Cammarato A, Li XE, Reedy MC, Lee CF, Lehman W, Bernstein SI. Structural basis for myopathic defects engendered by alterations in the myosin rod. J Mol Biol. 2011 Dec 9;414(4):477-84. doi: 10.1016/j.jmb.2011.10.019. Epub 2011 Oct 20. PubMed PMID: 22037585; PubMed Central PMCID: PMC3230674.
17: Wang Y, Melkani GC, Suggs JA, Melkani A, Kronert WA, Cammarato A*, Bernstein SI*. Expression of the inclusion body myopathy 3 mutation in Drosophila depresses myosin function and stability and recapitulates muscle inclusions and weakness. Mol Biol Cell. 2012 Jun;23(11):2057-65. doi: 10.1091/mbc.E12-02-0120. Epub 2012 Apr 11. PubMed PMID: 22496423; PubMed Central PMCID: PMC3364171. (* co-corresponding authors)
18: Xie HB, Cammarato A, Rajasekaran NS, Zhang H, Suggs JA, Lin HC, Bernstein SI, Benjamin IJ, Golic KG. The NADPH metabolic network regulates human αB-crystallin cardiomyopathy and reductive stress in Drosophila melanogaster. PLoS Genet. 2013 Jun;9(6):e1003544. doi: 10.1371/journal.pgen.1003544. Epub 2013 Jun 20. PubMed PMID: 23818860; PubMed Central PMCID: PMC3688542.
19: Viswanathan MC, Kaushik G, Engler AJ, Lehman W, Cammarato A*. A Drosophila melanogaster model of diastolic dysfunction and cardiomyopathy based on impaired troponin-T function. Circ Res. 2014 Jan 17;114(2):e6-17. doi: 10.1161/CIRCRESAHA.114.302028. Epub 2013 Nov 12. Erratum in: Circ Res. 2014 Feb 14;114(4):e28. PubMed PMID: 24221941; PubMed Central PMCID: PMC4526186. (* corresponding author)
20: Nishimura M, Kumsta C, Kaushik G, Diop SB, Ding Y, Bisharat-Kernizan J, Catan H, Cammarato A, Ross RS, Engler AJ, Bodmer R, Hansen M, Ocorr K. A dual role for integrin-linked kinase and β1-integrin in modulating cardiac aging. Aging Cell. 2014 Jun;13(3):431-40. doi: 10.1111/acel.12193. Epub 2014 Jan 9. PubMed PMID: 24400780; PubMed Central PMCID: PMC4032615.
21: Viswanathan MC, Blice-Baum AC, Schmidt W, Foster DB, Cammarato A*. Pseudo-acetylation of K326 and K328 of actin disrupts Drosophila melanogaster indirect flight muscle structure and performance. Front Physiol. 2015 Apr 28;6:116. doi: 10.3389/fphys.2015.00116. eCollection 2015. PubMed PMID: 25972811; PubMed Central PMCID: PMC4412121. (* corresponding author)
22: Cammarato A, Ocorr S, Ocorr K. Enhanced assessment of contractile dynamics in Drosophila hearts. Biotechniques. 2015 Feb 1;58(2):77-80. doi: 10.2144/000114255. eCollection 2015 Feb. PubMed PMID: 25652030.
23: Kaushik G, Spenlehauer A, Sessions AO, Trujillo AS, Fuhrmann A, Fu Z, Venkatraman V, Pohl D, Tuler J, Wang M, Lakatta EG, Ocorr K, Bodmer R, Bernstein SI, Van Eyk JE, Cammarato A*, Engler AJ*. Vinculin network-mediated cytoskeletal remodeling regulates contractile function in the aging heart. Sci Transl Med. 2015 Jun 17;7(292):292ra99. doi: 10.1126/scitranslmed.aaa5843. PubMed PMID: 26084806; PubMed Central PMCID: PMC4507505. (* co-corresponding authors)
24: Kooij V, Viswanathan MC, Lee DI, Rainer PP, Schmidt W, Kronert WA, Harding SE, Kass DA, Bernstein SI, Van Eyk JE, Cammarato A. Profilin modulates sarcomeric organization and mediates cardiomyocyte hypertrophy. Cardiovasc Res. 2016 May 15;110(2):238-48. doi: 10.1093/cvr/cvw050. Epub 2016 Mar 7. PubMed PMID: 26956799; PubMed Central PMCID: PMC4836629.
25: Viswanathan MC, Blice-Baum AC, Sang TK, Cammarato A*. Cardiac-Restricted Expression of VCP/TER94 RNAi or Disease Alleles Perturbs Drosophila Heart Structure and Impairs Function. J Cardiovasc Dev Dis. 2016 Jun;3(2). pii: 19. Epub 2016 May 24. PubMed PMID: 27500162; PubMed Central PMCID: PMC4973812. (* corresponding author)
26: Achal M, Trujillo AS, Melkani GC, Farman GP, Ocorr K, Viswanathan MC, Kaushik G, Newhard CS, Glasheen BM, Melkani A, Suggs JA, Moore JR, Swank DM, Bodmer R, Cammarato A, Bernstein SI. A restrictive cardiomyopathy mutation in an invariant proline at the myosin head/rod junction enhances head flexibility and function, yielding muscle defects in Drosophila. J Mol Biol. 2016 Jun 5;428(11):2446-61. doi: 10.1016/j.jmb.2016.04.021. Epub 2016 Apr 20. PubMed PMID: 27107639; PubMed Central PMCID: PMC4884507.
27: Blice-Baum AC, Zambon AC, Kaushik G, Viswanathan MC, Engler AJ, Bodmer R, Cammarato A*. Modest overexpression of FOXO maintains cardiac proteostasis and ameliorates age-associated functional decline. Aging Cell. 2016. In press. (* corresponding author)
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1: Taghli-Lamallem O*, Bodmer R, Chamberlain JS, Cammarato A*. Genetics and pathogenic mechanisms of cardiomyopathies in the Drosophila model. Drug Discovery Today: Disease Models. 2008 Nov 30;5(3):125-34. (* co-corresponding authors)
2: Kaushik G, Zambon AC, Fuhrmann A, Bernstein SI, Bodmer R, Engler AJ, Cammarato A*. Measuring passive myocardial stiffness in Drosophila melanogaster to investigate diastolic dysfunction. J Cell Mol Med. 2012 Aug;16(8):1656-62. doi: 10.1111/j.1582-4934.2011.01517.x. PubMed PMID: 22225769; PubMed Central PMCID: PMC3326184. (* corresponding author)
3: Kooij V, Venkatraman V, Tra J, Kirk JA, Rowell J, Blice-Baum AC, Cammarato A, Van Eyk JE. Sizing up models of heart failure: Proteomics from flies to humans. Proteomics Clin Appl. 2014 Oct;8(9-10):653-64. doi: 10.1002/prca.201300123. Epub 2014 Jun 25. PubMed PMID: 24723306; PubMed Central PMCID: PMC4282793.
Honors and Awards
- Research article entitled “The NADPH metabolic network regulates human αB-crystallin cardiomyopathy and reductive stress in Drosophila melanogaster” was selected as “Featured Research” by PLoS Genetics and as the “Biomedical Picture of the Day” by the MRC Clinical Sciences Centre
- Research article entitled “A mighty small heart: the cardiac proteome of adult Drosophila Melanogaster” was selected as a top advance in functional genomics and translational biology for 2011 by Circulation: Cardiovascular Genetics
- Circulation, American Heart Association
Top Advances in Functional Genomics and Translational Biology for 2011 - EveryONE, PLOS Blogs
Drosophila Research Captures our Hearts, and Attention - Research article entitled “Myosin transducer mutations differentially affect motor function, myofibril structure and the performance of skeletal and cardiac muscles” was featured in the ASCB Newsletter “InCytes from MBC”. “InCytes” highlights important research findings and advances in cell biology from articles published in the corresponding monthly issue of MBC
Myosin Transducer Mutation Differentially Affect Motor Function, Myogibril Structure, and the Performance of Skeletal and Cardiac Muscles
Anthony Cammarato, Corey M. Dambacher, Aileen F. Knowles, William A. Kronert, Rolf Bodmer, Karen Ocorr, and Sanford I. Bernstein
Drosophila, which have a single muscle myosin gene whose splice variants are expressed in every striated muscle, are an ideal model system for analyzing the pathological consequences of mutations that alter the motor's chemomechanical properties. The authors have studied two mutations located in the transducer domain of the muscle myosin motor, D45 and Mhc5, which, respectively, decrease and increase ATPase activity and motility in vitro. The hypoactive D45 mutant protects against age-associated dysfunction of metabolically demanding skeletral muscles but causes a dilated cardiomyopathy phenotype similar to that seen in human patients with hypoactive cardiac myosin mutations. In contrast, the hyperactive Mhc5 mutant disrupts the ulatrstructure and function of skeletal muscles, reflecting disinhibition and hypercontraction. The cardiac phenotype in Mhc5-expressing flies resembles a rare myocardial disorder, human restricive cardiomyopathy, that although not previously linked to myosin mutations is associated with dysregulation of motor activity. The authors' prediction that Drosophila may serve as a useful model in this regard has been borne out by very recent documentation that a myosin mutation causes pediatric restrictive cardiomyopathy. (S.M. Ware, et al., Clin. Genetic., in press).