DOCSLIB.ORG

Normalization of NAD+ Redox Balance As a Therapy for Heart Failure ORIGINAL RESEARCH ARTICLE

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Normalization of NAD+ Redox Balance As a Therapy for Heart Failure ORIGINAL RESEARCH ARTICLE ORIGINAL RESEARCH ARTICLE 883 Rong Tian, permeability hypertrophy hypertrophy ◼ ◼ cardiac metabolism cardiac September 20, 2016 September 20, DVM heart failure heart failure mitochondria Correspondence to: Correspondence MD, PhD, Mitochondria and University of Metabolism Center, Room N130, 850 Washington, Seattle, WA Republican Street, 98109-8057. E-mail rongtian@u. washington.edu of Funding, see page 893 Sources Key Words: ◼ ◼ transition pore © 2016 American Heart Association, Inc. Chi Fung Lee, PhD Juan D. Chavez, PhD Garcia-Menendez, Lorena Choi, MD Yongseon Nathan D. Roe, PhD Ann Chiao, PhD Ying PhD John S. Edgar, Ah Goo, PhD Young David R. Goodlett, PhD James E. Bruce, PhD Rong Tian, MD, PhD salvage + edox Balance ailure redox balance either redox ratio. Hyperacetylation + + R + D eart F RTICLE NA H imbalance in the failing hearts. These A ratio) and protein acetylation in the ratio) and protein and protein hyperacetylation, previously hyperacetylation, previously and protein + + + levels by stimulating the NAD + precursors are feasible in humans. are precursors ESEARCH + We show that mitochondrial protein hyperacetylation due show that mitochondrial protein We Impairments of mitochondrial function in the heart are Impairments of mitochondrial function in the heart are R herapy for We assessed the reduced/oxidized ratio of nicotinamide assessed the reduced/oxidized We Increased NADH/NAD Increased T redox imbalance contributes to the pathologic remodeling of the imbalance contributes to the pathologic remodeling redox 2016;134:883–894. DOI: 10.1161/CIRCULATIONAHA.116.022495 DOI: 2016;134:883–894. + pool also were tested. pool also were + ormalization of pathway suppressed mitochondrial protein hyperacetylation and cardiac hyperacetylation and cardiac mitochondrial protein pathway suppressed to stresses. function in responses cardiac and improved hypertrophy, Acetylome analysis identified a subpopulation of mitochondrial proteins that was sensitive to changes in the NADH/NAD Circulation. METHODS: adenine dinucleotide (NADH/NAD BACKGROUND: is no therapy but there linked intricately to the development of heart failure, for mitochondrial dysfunction. N ORIGINAL RESULTS: heart via 2 distinct mechanisms. Our preclinical data demonstrate a clear heart via 2 distinct mechanisms. Our preclinical benefit of normalizing NADH/NAD observed in genetic models of defective mitochondrial function, also are observed in genetic models of defective mitochondrial function, also are in human failing hearts as well as in mouse heartspresent with pathologic Elevation of NAD hypertrophy. failing heart. Proteome and acetylome analyses were followed by docking and acetylome analyses were failing heart. Proteome calculation, mutagenesis, and mitochondrial calcium uptake assays to of specific acetylation sites. The therapeutic determine the functional role acetylation by expanding the effects of normalizing mitochondrial protein NAD findings have a high translational potential as the pharmacologic strategy NAD of increasing of mitochondrial malate-aspartate shuttle proteins impaired the transport impaired of mitochondrial malate-aspartate shuttle proteins in altered and oxidation of cytosolic NADH in the mitochondria, resulting acetylation Furthermore, deficiency. state and energy cytosolic redox at lysine-70 in adenosine of oligomycin-sensitive conferring protein its interaction with cyclophilin D, triphosphate synthase complex promoted and sensitized the opening of mitochondrial permeability transition pore. Both could be alleviated by normalizing the NAD CONCLUSIONS: to NAD genetically or pharmacologically. as a Downloaded from http://circ.ahajournals.org/ by guest on October 6, 2016 Lee et al transferase, the later is determined by the deacetylase Clinical Perspective activity, and primarily sirtuins in the mitochondria. The sirtuin deacetylases consume NAD+ as a cosubstrate9; What Is New? mitochondrial function is critical for setting the NADH/ + + • We show that mitochondrial dysfunction increases NAD balance thus the NAD available for sirtuin activity. NADH/NAD+ ratio and protein hyperacetylation result- Using a mouse model with primary mitochondrial dys- ing in a greater sensitivity of the heart to chronic stress. function (cardiac-specific deletion of a Complex-I protein, • Mechanistically, we have identified that hyperacety- Ndufs4: cKO), we recently found that elevation in NADH/ lation of the regulators of mitochondrial permeability NAD+ ratio induce mitochondrial protein hyperacetylation transition pore and malate aspartate shuttle medi- and renders the heart highly susceptible to stresses.10 In ates the increased susceptibility to stresses. this study we defined the molecular intermediaries link- • Increased acetylation levels of cyclophilin D and ing specific NAD+-sensitive hyperacetylation targets to oligomycin sensitive conferring protein on the ade- the development of heart failure and demonstrated the nosine triphosphate synthase complex promote the interaction between oligomycin sensitive conferring relevance of these mechanisms in human heart failure. + protein and cyclophilin D and increase mitochondrial Furthermore, we showed that restoring the NADH/NAD permeability transition pore sensitivity. Moreover, ratio by genetic and pharmacologic approaches is an ef- hyperacetylation and inhibition of malate aspartate fective and potentially translatable strategy for the treat- Downloaded from shuttle limits the mitochondrial import and oxidation ment of heart failure in clinical practice. of NADH generated in the cytosol resulting in cyto- solic redox imbalance. METHODS What are the Clinical Implications? Animal Care, Surgical Procedures, and http://circ.ahajournals.org/ • The identified mechanisms described above are Echocardiography observed in animal models and human failing hearts. All procedures involving animal use were performed with the • Our preclinical results show that expanding the cardiac approval of Institutional Animal Care and Use Committee of NAD+ pool via pharmacologic or genetic approaches the University of Washington. Procedures for animal care, normalizes the NADH/NAD+ ratio and protein acetyla- surgeries, and echocardiography were in the online-only Data tion in hypertrophied and failing hearts. Supplement. • Importantly, these measures improve cardiac func- tion and reduce pathologic hypertrophy in mice. + by guest on October 6, 2016 Thus, the study identifies NADH/NAD ratio a viable Ex Vivo Measurements of Cardiac Function and therapeutic target for mitochondrial dysfunction and Energetics heart failure. Langendorff perfused mouse hearts were isolated as described in the online-only Data Supplement.11 ardiovascular disease is a leading cause of death Mitochondrial Isolation, Proteome, and worldwide.1 As the life expectancy increases and the mortality of acute ischemic events decreases, Acetylome Analyses C 12 the incidence of heart failure is mounting at a pace of Mitochondria were isolated as described. Peptide generation and mass spectrometric protocols were in the online-only Data 900 000 per year.2 However, medical therapy for heart Supplement. failure has been stalled for almost 2 decades. Novel con- Acquired tandem mass spectra were searched for sequence cepts and strategies in the treatment of heart failure are matches against the International Protein Index mouse database needed urgently. using SEQUEST. The following modifications were set as search The heart is a high energy-consuming organ. Mi- parameters: peptide mass tolerance at 500 ppm, trypsin diges- tochondrion is the powerhouse of the cell and mito- tion cleavage after K or R (except when followed by P), one chondrial dysfunction is a well-recognized maladaptive allowed missed cleavage site, carboxymethylated cysteines mechanism during the development of heart failure.3,4 (static modification), and oxidized methionines or acetylation on Targeting mitochondria for heart failure therapy has long K (variable modification). PeptideProphet13 and ProteinProphet14 been sought; however, previous work focusing on im- were used to assign confidence in the identified spectra result- proving mitochondrial energy production and reducing ing from the SEQUEST search. It relies on probability models reactive oxygen species yielded few successful clinical and an empirical Bayesian approach to model fitting. First a score is produced to reflect the quality of each spectrum. Then applications.5 In recent years, protein lysine acetylation a probability-based model is produced for the distribution of emerged as an important mechanism linking mitochon- correctly and incorrectly identified spectra and fit to the scores 6–8 drial metabolism to cellular pathologies. The level of of all identified spectra. The confidence in individual spectra protein acetylation reflects the balance of acetylation are evaluated using the posterior probability. A cutoff is applied and deacetylation. Although the former is dependent on on the scores for the set of correctly identified spectra to the abundance of acetyl-CoA and the activity of acetyl- control the false discovery rate, defined as the percentage of 884 September 20, 2016 Circulation. 2016;134:883–894. DOI: 10.1161/CIRCULATIONAHA.116.022495 Normalizing NADH/NAD+ as Heart Failure Therapy false positives which pass the cutoff. This method produces a caused by mitochondrial Complex-I deficiency (cKO) in- similar estimation of the false discovery rate as the commonly creased cardiac susceptibility to stress.10 We sought used
Load more

Recommended publications
  • The Mitochondrial Kinase PINK1 in Diabetic Kidney Disease
    International Journal of Molecular Sciences Review The Mitochondrial Kinase PINK1 in Diabetic Kidney Disease Chunling Huang * , Ji Bian , Qinghua Cao, Xin-Ming Chen and Carol A. Pollock * Kolling Institute, Sydney Medical School, Royal North Shore Hospital, University of Sydney, St. Leonards, NSW 2065, Australia; [email protected] (J.B.); [email protected] (Q.C.); [email protected] (X.-M.C.) * Correspondence: [email protected] (C.H.); [email protected] (C.A.P.); Tel.: +61-2-9926-4784 (C.H.); +61-2-9926-4652 (C.A.P.) Abstract: Mitochondria are critical organelles that play a key role in cellular metabolism, survival, and homeostasis. Mitochondrial dysfunction has been implicated in the pathogenesis of diabetic kidney disease. The function of mitochondria is critically regulated by several mitochondrial protein kinases, including the phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1). The focus of PINK1 research has been centered on neuronal diseases. Recent studies have revealed a close link between PINK1 and many other diseases including kidney diseases. This review will provide a concise summary of PINK1 and its regulation of mitochondrial function in health and disease. The physiological role of PINK1 in the major cells involved in diabetic kidney disease including proximal tubular cells and podocytes will also be summarized. Collectively, these studies suggested that targeting PINK1 may offer a promising alternative for the treatment of diabetic kidney disease. Keywords: PINK1; diabetic kidney disease; mitochondria; mitochondria quality control; mitophagy Citation: Huang, C.; Bian, J.; Cao, Q.; 1. Introduction Chen, X.-M.; Pollock, C.A.
    [Show full text]
  • Molecular Genetic Delineation of 2Q37.3 Deletion in Autism and Osteodystrophy: Report of a Case and of New Markers for Deletion Screening by PCR
    UC Irvine UC Irvine Previously Published Works Title Molecular genetic delineation of 2q37.3 deletion in autism and osteodystrophy: report of a case and of new markers for deletion screening by PCR. Permalink https://escholarship.org/uc/item/83f0x61r Journal Cytogenetics and cell genetics, 94(1-2) ISSN 0301-0171 Authors Smith, M Escamilla, JR Filipek, P et al. Publication Date 2001 DOI 10.1159/000048775 License https://creativecommons.org/licenses/by/4.0/ 4.0 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Original Article Cytogenet Cell Genet 94:15–22 (2001) Molecular genetic delineation of 2q37.3 deletion in autism and osteodystrophy: report of a case and of new markers for deletion screening by PCR M. Smith, J.R. Escamilla, P. Filipek, M.E. Bocian, C. Modahl, P. Flodman, and M.A. Spence Department of Pediatrics, University of California, Irvine CA (USA) Abstract. We recently studied a patient who meets criteria us to determine the parental origin of the deletion in our for autistic disorder and has a 2q37 deletion. Molecular cyto- patient. DNA from 8–13 unrelated individuals was used to genetic studies were carried out using DNA isolated from 22 determine heterozygosity estimates for these markers. We re- different 2q37 mapped BACs to more precisely define the view four genes deleted in our patient – genes whose known extent of the chromosome deletion. We also analyzed 2q37 functions and sites of expression in the brain and/or bone make mapped polymorphic markers. In addition DNA sequences of them candidates for involvement in autism and/or the osteo- BACs in the deletion region were scanned to identify microsa- dystrophy observed in patients with 2q37.3 deletions.
    [Show full text]
  • Proteomic and Metabolomic Analyses of Mitochondrial Complex I-Deficient
    THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 287, NO. 24, pp. 20652–20663, June 8, 2012 © 2012 by The American Society for Biochemistry and Molecular Biology, Inc. Published in the U.S.A. Proteomic and Metabolomic Analyses of Mitochondrial Complex I-deficient Mouse Model Generated by Spontaneous B2 Short Interspersed Nuclear Element (SINE) Insertion into NADH Dehydrogenase (Ubiquinone) Fe-S Protein 4 (Ndufs4) Gene*□S Received for publication, November 25, 2011, and in revised form, April 5, 2012 Published, JBC Papers in Press, April 25, 2012, DOI 10.1074/jbc.M111.327601 Dillon W. Leong,a1 Jasper C. Komen,b1 Chelsee A. Hewitt,a Estelle Arnaud,c Matthew McKenzie,d Belinda Phipson,e Melanie Bahlo,e,f Adrienne Laskowski,b Sarah A. Kinkel,a,g,h Gayle M. Davey,g William R. Heath,g Anne K. Voss,a,h René P. Zahedi,i James J. Pitt,j Roman Chrast,c Albert Sickmann,i,k Michael T. Ryan,l Gordon K. Smyth,e,f,h b2 a,h,m,n3 David R. Thorburn, and Hamish S. Scott Downloaded from From the aMolecular Medicine Division, gImmunology Division, and eBioinformatics Division, Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia, the bMurdoch Childrens Research Institute, Royal Children’s Hospital and Department of Paediatrics, University of Melbourne, Parkville, Victoria 3052, Australia, the cDépartement de Génétique Médicale, Université de Lausanne, 1005 Lausanne, Switzerland, the dCentre for Reproduction and Development, Monash Institute of Medical Research, Clayton, Victoria 3168, Australia, the hDepartment of Medical Biology
    [Show full text]
  • Low Abundance of the Matrix Arm of Complex I in Mitochondria Predicts Longevity in Mice
    ARTICLE Received 24 Jan 2014 | Accepted 9 Apr 2014 | Published 12 May 2014 DOI: 10.1038/ncomms4837 OPEN Low abundance of the matrix arm of complex I in mitochondria predicts longevity in mice Satomi Miwa1, Howsun Jow2, Karen Baty3, Amy Johnson1, Rafal Czapiewski1, Gabriele Saretzki1, Achim Treumann3 & Thomas von Zglinicki1 Mitochondrial function is an important determinant of the ageing process; however, the mitochondrial properties that enable longevity are not well understood. Here we show that optimal assembly of mitochondrial complex I predicts longevity in mice. Using an unbiased high-coverage high-confidence approach, we demonstrate that electron transport chain proteins, especially the matrix arm subunits of complex I, are decreased in young long-living mice, which is associated with improved complex I assembly, higher complex I-linked state 3 oxygen consumption rates and decreased superoxide production, whereas the opposite is seen in old mice. Disruption of complex I assembly reduces oxidative metabolism with concomitant increase in mitochondrial superoxide production. This is rescued by knockdown of the mitochondrial chaperone, prohibitin. Disrupted complex I assembly causes premature senescence in primary cells. We propose that lower abundance of free catalytic complex I components supports complex I assembly, efficacy of substrate utilization and minimal ROS production, enabling enhanced longevity. 1 Institute for Ageing and Health, Newcastle University, Newcastle upon Tyne NE4 5PL, UK. 2 Centre for Integrated Systems Biology of Ageing and Nutrition, Newcastle University, Newcastle upon Tyne NE4 5PL, UK. 3 Newcastle University Protein and Proteome Analysis, Devonshire Building, Devonshire Terrace, Newcastle upon Tyne NE1 7RU, UK. Correspondence and requests for materials should be addressed to T.v.Z.
    [Show full text]
  • Mitoxplorer, a Visual Data Mining Platform To
    mitoXplorer, a visual data mining platform to systematically analyze and visualize mitochondrial expression dynamics and mutations Annie Yim, Prasanna Koti, Adrien Bonnard, Fabio Marchiano, Milena Dürrbaum, Cecilia Garcia-Perez, José Villaveces, Salma Gamal, Giovanni Cardone, Fabiana Perocchi, et al. To cite this version: Annie Yim, Prasanna Koti, Adrien Bonnard, Fabio Marchiano, Milena Dürrbaum, et al.. mitoXplorer, a visual data mining platform to systematically analyze and visualize mitochondrial expression dy- namics and mutations. Nucleic Acids Research, Oxford University Press, 2020, 10.1093/nar/gkz1128. hal-02394433 HAL Id: hal-02394433 https://hal-amu.archives-ouvertes.fr/hal-02394433 Submitted on 4 Dec 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution| 4.0 International License Nucleic Acids Research, 2019 1 doi: 10.1093/nar/gkz1128 Downloaded from https://academic.oup.com/nar/advance-article-abstract/doi/10.1093/nar/gkz1128/5651332 by Bibliothèque de l'université la Méditerranée user on 04 December 2019 mitoXplorer, a visual data mining platform to systematically analyze and visualize mitochondrial expression dynamics and mutations Annie Yim1,†, Prasanna Koti1,†, Adrien Bonnard2, Fabio Marchiano3, Milena Durrbaum¨ 1, Cecilia Garcia-Perez4, Jose Villaveces1, Salma Gamal1, Giovanni Cardone1, Fabiana Perocchi4, Zuzana Storchova1,5 and Bianca H.
    [Show full text]
  • A CRISPR-Cas9 Screen Identifies Mitochondrial Translation As An
    A CRISPR-Cas9 screen identifies mitochondrial translation as an essential process in latent KSHV infection of human endothelial cells Daniel L. Holmesa , Daniel T. Vogta , and Michael Lagunoffa,1 aDepartment of Microbiology, University of Washington, Seattle, WA 98109 Edited by Donald E. Ganem, Novartis Institutes for Biomedical Research, Inc., Emeryville, CA, and approved September 30, 2020 (received for review June 9, 2020) Kaposi’s sarcoma-associated herpesvirus (KSHV) is the etiologic KS tumors, providing a culture model for KSHV latency in agent of Kaposi’s sarcoma (KS) and primary effusion lymphoma tumors (3). Using cell culture systems, our laboratory has iden- (PEL). The main proliferating component of KS tumors is a tified several cellular pathways, which can be used to selec- cell of endothelial origin termed the spindle cell. Spindle cells tively target latently infected cells in vitro (4–7). Recently are predominantly latently infected with only a small percent- developed lentivirus-encoded CRISPR-Cas9–based screening age of cells undergoing viral replication. As there is no direct platforms have enabled large-scale interrogation of so-called treatment for latent KSHV, identification of host vulnerabili- “Achilles” genes within a population of human cells (8). These ties in latently infected endothelial cells could be exploited screens can be used to identify factors that are essential to the to inhibit KSHV-associated tumor cells. Using a pooled CRISPR- survival of cancer cells. Similar approaches have been used to Cas9 lentivirus library, we identified host factors that are identify genes that are critical for the survival of B cell lym- essential for the survival or proliferation of latently infected phomas infected with Epstein-Barr virus (EBV) (9).
    [Show full text]
  • Human CLPB) Is a Potent Mitochondrial Protein Disaggregase That Is Inactivated By
    bioRxiv preprint doi: https://doi.org/10.1101/2020.01.17.911016; this version posted January 18, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Skd3 (human CLPB) is a potent mitochondrial protein disaggregase that is inactivated by 3-methylglutaconic aciduria-linked mutations Ryan R. Cupo1,2 and James Shorter1,2* 1Department of Biochemistry and Biophysics, 2Pharmacology Graduate Group, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, U.S.A. *Correspondence: [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.01.17.911016; this version posted January 18, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. ABSTRACT Cells have evolved specialized protein disaggregases to reverse toxic protein aggregation and restore protein functionality. In nonmetazoan eukaryotes, the AAA+ disaggregase Hsp78 resolubilizes and reactivates proteins in mitochondria. Curiously, metazoa lack Hsp78. Hence, whether metazoan mitochondria reactivate aggregated proteins is unknown. Here, we establish that a mitochondrial AAA+ protein, Skd3 (human CLPB), couples ATP hydrolysis to protein disaggregation and reactivation. The Skd3 ankyrin-repeat domain combines with conserved AAA+ elements to enable stand-alone disaggregase activity. A mitochondrial inner-membrane protease, PARL, removes an autoinhibitory peptide from Skd3 to greatly enhance disaggregase activity. Indeed, PARL-activated Skd3 dissolves α-synuclein fibrils connected to Parkinson’s disease. Human cells lacking Skd3 exhibit reduced solubility of various mitochondrial proteins, including anti-apoptotic Hax1.
    [Show full text]
  • Skd3 (Human CLPB) Is a Potent Mitochondrial Protein Disaggregase That Is Inactivated By
    bioRxiv preprint first posted online Jan. 18, 2020; doi: http://dx.doi.org/10.1101/2020.01.17.911016. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. Skd3 (human CLPB) is a potent mitochondrial protein disaggregase that is inactivated by 3-methylglutaconic aciduria-linked mutations Ryan R. Cupo1,2 and James Shorter1,2* 1Department of Biochemistry and Biophysics, 2Pharmacology Graduate Group, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, U.S.A. *Correspondence: [email protected] 1 bioRxiv preprint first posted online Jan. 18, 2020; doi: http://dx.doi.org/10.1101/2020.01.17.911016. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. ABSTRACT Cells have evolved specialized protein disaggregases to reverse toxic protein aggregation and restore protein functionality. In nonmetazoan eukaryotes, the AAA+ disaggregase Hsp78 resolubilizes and reactivates proteins in mitochondria. Curiously, metazoa lack Hsp78. Hence, whether metazoan mitochondria reactivate aggregated proteins is unknown. Here, we establish that a mitochondrial AAA+ protein, Skd3 (human CLPB), couples ATP hydrolysis to protein disaggregation and reactivation. The Skd3 ankyrin-repeat domain combines with conserved AAA+ elements to enable stand-alone disaggregase activity. A mitochondrial inner-membrane protease, PARL, removes an autoinhibitory peptide from Skd3 to greatly enhance disaggregase activity. Indeed, PARL-activated Skd3 dissolves α-synuclein fibrils connected to Parkinson’s disease.
    [Show full text]
  • THE FUNCTIONAL SIGNIFICANCE of MITOCHONDRIAL SUPERCOMPLEXES in C. ELEGANS by WICHIT SUTHAMMARAK Submitted in Partial Fulfillment
    THE FUNCTIONAL SIGNIFICANCE OF MITOCHONDRIAL SUPERCOMPLEXES in C. ELEGANS by WICHIT SUTHAMMARAK Submitted in partial fulfillment of the requirements For the degree of Doctor of Philosophy Dissertation Advisor: Drs. Margaret M. Sedensky & Philip G. Morgan Department of Genetics CASE WESTERN RESERVE UNIVERSITY January, 2011 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the thesis/dissertation of _____________________________________________________ candidate for the ______________________degree *. (signed)_______________________________________________ (chair of the committee) ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ (date) _______________________ *We also certify that written approval has been obtained for any proprietary material contained therein. Dedicated to my family, my teachers and all of my beloved ones for their love and support ii ACKNOWLEDGEMENTS My advanced academic journey began 5 years ago on the opposite side of the world. I traveled to the United States from Thailand in search of a better understanding of science so that one day I can return to my homeland and apply the knowledge and experience I have gained to improve the lives of those affected by sickness and disease yet unanswered by science. Ultimately, I hoped to make the academic transition into the scholarly community by proving myself through scientific research and understanding so that I can make a meaningful contribution to both the scientific and medical communities. The following dissertation would not have been possible without the help, support, and guidance of a lot of people both near and far. I wish to thank all who have aided me in one way or another on this long yet rewarding journey. My sincerest thanks and appreciation goes to my advisors Philip Morgan and Margaret Sedensky.
    [Show full text]
  • Table 3: Average Gene Expression Profiles by Chromosome
    Supplemental Data Table 1: Experimental Setup Correlation Array Reverse Fluor Array Extraction Coefficient Print Batch (Y/N) mean (range) DLD1-I.1 I A N DLD1-I.2 I B N 0.86 DLD1-I.3 I C N (0.79-0.90) DLD1-I.4 I C Y DLD1 DLD1-II.1 II D N DLD1-II.2 II E N 0.86 DLD1-II.3 II F N (0.74-0.94) DLD1-II.4 II F Y DLD1+3-II.1 II A N DLD1+3-II.2 II A N 0.85 DLD1 + 3 DLD1+3-II.3 II B N (0.64-0.95) DLD1+3-II.4 II B Y DLD1+7-I.1 I A N DLD1+7-I.2 I A N 0.79 DLD1 + 7 DLD1+7-I.3 I B N (0.68-0.90) DLD1+7-I.4 I B Y DLD1+13-I.1 I A N DLD1+13-I.2 I A N 0.88 DLD1 + 13 DLD1+13-I.3 I B N (0.84-0.91) DLD1+13-I.4 I B Y hTERT-HME-I.1 I A N hTERT-HME-I.2 I B N 0.85 hTERT-HME hTERT-HME-I.3 I C N (0.80-0.92) hTERT-HME-I.4 I C Y hTERT-HME+3-I.1 I A N hTERT-HME+3-I.2 I B N 0.84 hTERT-HME + 3 hTERT-HME+3-I.3 I C N (0.74-0.90) hTERT-HME+3-I.4 I C Y Supplemental Data Table 2: Average gene expression profiles by chromosome arm DLD1 hTERT-HME Ratio.7 Ratio.1 Ratio.3 Ratio.3 Chrom.
    [Show full text]
  • Influenza-Specific Effector Memory B Cells Predict Long-Lived Antibody Responses to Vaccination in Humans
    bioRxiv preprint doi: https://doi.org/10.1101/643973; this version posted February 18, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Influenza-specific effector memory B cells predict long-lived antibody responses to vaccination in humans Anoma Nellore1, Esther Zumaquero2, Christopher D. Scharer3, Rodney G. King2, Christopher M. Tipton4, Christopher F. Fucile5, Tian Mi3, Betty Mousseau2, John E. Bradley6, Fen Zhou2, Paul A. Goepfert1, Jeremy M. Boss3, Troy D. Randall6, Ignacio Sanz4, Alexander F. Rosenberg2,5, Frances E. Lund2 1Dept. of Medicine, Division of Infectious Disease, 2Dept of Microbiology, 5Informatics Institute, 6Dept. of Medicine, Division of Clinical Immunology and Rheumatology and at The University of Alabama at Birmingham, Birmingham, AL 35294 USA 3Dept. of Microbiology and Immunology and 4Department of Medicine, Division of Rheumatology Emory University, Atlanta, GA 30322, USA Correspondence should be addressed to: Frances E. Lund, PhD Charles H. McCauley Professor and Chair Dept of Microbiology University of Alabama at Birmingham 276 BBRB Box 11 1720 2nd Avenue South Birmingham AL 35294-2170 [email protected] SHORT RUNNING TITLE: Effector memory B cell development after influenza vaccination 1 bioRxiv preprint doi: https://doi.org/10.1101/643973; this version posted February 18, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract Seasonal influenza vaccination elicits hemagglutinin (HA)-specific CD27+ memory B cells (Bmem) that differ in expression of T-bet, BACH2 and TCF7.
    [Show full text]
  • Electron Transport Chain Activity Is a Predictor and Target for Venetoclax Sensitivity in Multiple Myeloma
    ARTICLE https://doi.org/10.1038/s41467-020-15051-z OPEN Electron transport chain activity is a predictor and target for venetoclax sensitivity in multiple myeloma Richa Bajpai1,7, Aditi Sharma 1,7, Abhinav Achreja2,3, Claudia L. Edgar1, Changyong Wei1, Arusha A. Siddiqa1, Vikas A. Gupta1, Shannon M. Matulis1, Samuel K. McBrayer 4, Anjali Mittal3,5, Manali Rupji 6, Benjamin G. Barwick 1, Sagar Lonial1, Ajay K. Nooka 1, Lawrence H. Boise 1, Deepak Nagrath2,3,5 & ✉ Mala Shanmugam 1 1234567890():,; The BCL-2 antagonist venetoclax is highly effective in multiple myeloma (MM) patients exhibiting the 11;14 translocation, the mechanistic basis of which is unknown. In evaluating cellular energetics and metabolism of t(11;14) and non-t(11;14) MM, we determine that venetoclax-sensitive myeloma has reduced mitochondrial respiration. Consistent with this, low electron transport chain (ETC) Complex I and Complex II activities correlate with venetoclax sensitivity. Inhibition of Complex I, using IACS-010759, an orally bioavailable Complex I inhibitor in clinical trials, as well as succinate ubiquinone reductase (SQR) activity of Complex II, using thenoyltrifluoroacetone (TTFA) or introduction of SDHC R72C mutant, independently sensitize resistant MM to venetoclax. We demonstrate that ETC inhibition increases BCL-2 dependence and the ‘primed’ state via the ATF4-BIM/NOXA axis. Further, SQR activity correlates with venetoclax sensitivity in patient samples irrespective of t(11;14) status. Use of SQR activity in a functional-biomarker informed manner may better select for MM patients responsive to venetoclax therapy. 1 Department of Hematology and Medical Oncology, Winship Cancer Institute, School of Medicine, Emory University, Atlanta, GA, USA.
    [Show full text]