J. Marie Hardwick
Our laboratory studies the basic molecular mechanisms of programmed cell death, an evolutionarily conserved process to eliminate cells. Because these pathways normally contribute to the millions of cell deaths that occur per day per individual, defects in cell death underlie the range of human disorders from cancer (insufficient cell death) to neurological diseases (excessive death). We study these processes in the nervous system, in cancer models and during virus infection using mouse models, yeast genetics and biochemical approaches. We have shown that viruses can trigger cells to activate programmed cell death (Levine et al., Nature, 1993), and that both viral and cellular regulators of apoptosis, such as Bcl-2 family proteins and many other factors, can alter the outcome of a virus infection (Lewis et al., Nature Med, 1999). We seek the mechanisms that explain why Sindbis virus induces neuronal cell death in young animals, but fails to activate the death pathway in neurons of adult brains and in mosquitoes that transmit the virus in nature. Interestingly, cell death factors can also be cell survival factors (Cheng et al., Science, 1997), leading us to pursue their important alternative functions, or ‘day-jobs’, such as regulating mitochondrial fission and bioenergetics, synaptic activity in neurons, or nutrient-sensing in yeast. Although it remains controversial as to whether or not unicellular organisms are capable of undergoing programmed cell death, we have new compelling evidence that these pathways are evolutionarily conserved. Therefore, we have launched an effort to apply the genetic and proteomic tools available for yeast to model the death and survival mechanisms that are conserved between yeast and mammals.
Host cell death responses determine pathogenesis. Viruses manipulate the host, and host responses contribute to cell death and viral pathogenicity, including neurons of the brain. Cellular caspase proteases convert host, but not viral, anti-apoptotic proteins into cell death factors. Ongoing work investigates cell death regulation by the virus that causes COVID-19.
Levine B, Huang Q, Isaacs JT, Reed JC, Griffin DE, Hardwick JM. Conversion of lytic to persistent alphavirus infection by the bcl-2 cellular oncogene. Nature. 1993;361:739-42. https://pubmed.ncbi.nlm.nih.gov/8441470/ GS Cited >500 times
Cheng EH, Kirsch DG, Clem RJ, Ravi R, Kastan MB, Bedi A, et al. Conversion of Bcl-2 to a Bax-like death effector by caspases. Science. 1997;278:1966-8. https://pubmed.ncbi.nlm.nih.gov/9395403/ GS Citations >1000
Lewis J, Oyler GA, Ueno K, Fannjiang YR, Chau BN, Vornov J, Korsmeyer SJ, Zou S, Hardwick JM. Inhibition of virus-induced neuronal apoptosis by Bax. Nat Med. 1999;5:832-5. https://pubmed.ncbi.nlm.nih.gov/10395331/ GS Citations >100
Ivanovska I, Hardwick JM. Viruses activate a genetically conserved cell death pathway in a unicellular organism. J Cell Biol. 2005;170:391-9. https://pubmed.ncbi.nlm.nih.gov/16061692/ Featured in Editor’s Choice (same issue); and in Science STKE 296: p287. Citations >100
Ofengeim D, Chen YB, Miyawaki T, Li H, Sacchetti S, Flannery RJ, et al. N-terminally cleaved Bcl-xL mediates ischemia-induced neuronal death. Nat Neurosci. 2012;15:574-80 *Co-corr. https://pubmed.ncbi.nlm.nih.gov/22366758/ Featured in News & Views (same issue), and The Scientist. Citations >50 Navratil et al.
The severe acute respiratory syndrome coronavirus (SARS-CoV-2) envelope (E) protein harbors a conserved BH3-like sequence BioRxiv April 2020 https://www.biorxiv.org/content/10.1101/2020.04.09.033522v2 “Day-jobs” of cell death factors also contribute to cell fate. Perhaps all cell death regulators have critical roles in healthy cells that are poorly understood. We study these roles in mitochondria, neurons, tumor cells and during infection using microscopy, molecular biology, and genomic approaches applied to mammalian cells, mice, flies, and yeast.
Cheng EH, Levine B, Boise LH, Thompson CB, Hardwick JM. Bax-independent inhibition of apoptosis by Bcl-XL. Nature. 1996;379:554-6. https://pubmed.ncbi.nlm.nih.gov/8596636/ GS Citations >500
Fannjiang Y, Kim CH, Huganir RL, Zou S, Lindsten T, Thompson CB, Mito T, Traystman RJ, Larsen T, Griffin DE, Mandir AS, Dawson TM, Dike S, Sappington AL, Kerr DA, Jonas EA, Kaczmarek LK, Hardwick JM. BAK alters neuronal excitability and can switch from anti- to pro-death function during postnatal development. Dev Cell. 2003;4:575-85. https://pubmed.ncbi.nlm.nih.gov/12689595/ GS Citations >100
Berman SB, Chen YB, Qi B, McCaffery JM, Rucker EB, 3rd, Goebbels S, Nave KA, Arnold BA, Jonas EA, Pineda FJ, Hardwick JM. Bcl-x L increases mitochondrial fission, fusion, and biomass in neurons. J Cell Biol. 2009;184:707-19. https://pubmed.ncbi.nlm.nih.gov/19255249/ Cover list. Citations >100
Chen YB, Aon MA, Hsu YT, Soane L, Teng X, McCaffery JM, Cheng WC, Qi B, Li H, Alavian KN, Dayhoff-Brannigan M, Zou S, Pineda FJ, O’Rourke B, Ko YH, Pedersen PL, Kaczmarek LK, Jonas EA, Hardwick JM. Bcl-xL regulates mitochondrial energetics by stabilizing the inner membrane potential. J Cell Biol. 2011;195:263-76. https://pubmed.ncbi.nlm.nih.gov/21987637/ Citations >100
Tang HL, Tang HM, Fung MC, Hardwick JM. In vivo CaspaseTracker biosensor system for detecting anastasis and non-apoptotic caspase activity. Sci Rep. 2015;5:9015. https://www.ncbi.nlm.nih.gov/pubmed/25757939 GS Citations >50
Lamb HM, Hardwick JM. The Dark Side of Estrogen Stops Translation to Induce Apoptosis. Mol Cell. 2019;75:1087-9. https://www.ncbi.nlm.nih.gov/pubmed/31539505
Cell death mechanisms in fungi. Programmed cell death was originally assumed to exist only in animals and plants, and not in microbial species (fungi, parasites, bacteria). We developed methods to study cell death in human fungal pathogens – silent killers (>half million deaths per year worldwide).
Fannjiang Y, Cheng WC, Lee SJ, Qi B, Pevsner J, McCaffery JM, Hill RB, Basanez G, Hardwick JM. Mitochondrial fission proteins regulate programmed cell death in yeast. Genes Dev. 2004;18:2785-97. https://www.ncbi.nlm.nih.gov/pubmed/15520274 Citations >100
Teng X, Hardwick JM. Reliable method for detection of programmed cell death in yeast. Methods Mol Biol. 2009;559:335-42. https://www.ncbi.nlm.nih.gov/pubmed/19609767
Teng X, Cheng WC, Qi B, Yu TX, Ramachandran K, Boersma MD, Hattier T, Lehmann PV, Pineda FJ, Hardwick JM. Gene-dependent cell death in yeast. Cell Death Dis. 2011;2:e188. https://www.ncbi.nlm.nih.gov/pubmed/21814286
Hardwick JM. Do Fungi Undergo Apoptosis-Like Programmed Cell Death? mBio. 2018;9. https://www.ncbi.nlm.nih.gov/pubmed/30065087
Aouacheria A, Cunningham KW, Hardwick* JM, Palkova Z, Powers T, Severin FF, Vachova L. Comment on “Sterilizing immunity in the lung relies on targeting fungal apoptosis-like programmed cell death”. Science. 2018;360. https://www.ncbi.nlm.nih.gov/pubmed/29930109
ulkarni M, Stolp ZD, Hardwick JM. Targeting intrinsic cell death pathways to control fungal pathogens. Biochem Pharmacol. 2019;162:71-8. https://www.ncbi.nlm.nih.gov/pubmed/30660496
Neurodevelopmental disorders – underlying mechanisms related to nutrient sensing and genome evolution first identifiec tin yeast cell death studies now being pursued in mice. Studying cell death, genome evolution, and amino acid-sensing/TORC1 pathways in yeast uncovered functions for understudied KCTD family proteins in neurodevelopment.
Cheng WC, Teng X, Park HK, Tucker CM, Dunham MJ, Hardwick JM. Fis1 deficiency selects for compensatory mutations responsible for cell death and growth control defects. Cell Death Differ. 2008;15:1838-46. https://www.ncbi.nlm.nih.gov/pubmed/18756280 GS Citations >50
Teng X, Dayhoff-Brannigan M, Cheng WC, Gilbert CE, Sing CN, Diny NL, Wheelan SJ, Dunham MJ, Boeke JD, Pineda FJ, Hardwick JM. Genome-wide consequences of deleting any single gene. Mol Cell. 2013;52:485-94. https://www.ncbi.nlm.nih.gov/pubmed/24211263 Featured in The Scientist, “One gene, two mutations”; SGD New & Noteworthy, “Gene knockouts may not be so clean after all”; NIH/ NIGMS Biomedical Beat blo. GS Citations >100
Metz KA, Teng X, Coppens I, Lamb HM, Wagner BE, Rosenfeld JA, Chen X, Zhang Y, Kim HJ, Meadow ME, Wang TS, Haberlandt ED, Anderson GW, Leshinsky-Silver E, Bi W, Markello TC, Pratt M, Makhseed N, Garnica A, Danylchuk NR, Burrow TA, Jayakar P, McKnight D, Agadi S, Gbedawo H, Stanley C, Alber M, Prehl I, Peariso K, Ong MT, Mordekar SR, Parker MJ, Crooks D, Agrawal PB, Berry GT, Loddenkemper T, Yang Y, Maegawa GHB, Aouacheria A, Markle JG, Wohlschlegel JA, Hartman AL, Hardwick JM. KCTD7 deficiency defines a distinct neurodegenerative disorder with a conserved autophagy-lysosome defect. Ann Neurol. 2018;84:766-80. https://www.ncbi.nlm.nih.gov/pubmed/30295347 Cover article.
Chen X, Wang G, Zhang Y, Dayhoff-Brannigan M, Diny NL, Zhao M, He G, Sing CN, Metz KA, Stolp ZD, Aouacheria A, Cheng WC, Hardwick JM, Teng X. Whi2 is a conserved negative regulator of TORC1 in response to low amino acids. PLoS Genet. 2018;14:e1007592. https://www.ncbi.nlm.nih.gov/pubmed/30142151
Teng X, Aouacheria A, Lionnard L, Metz KA, Soane L, Kamiya A, Hardwick JM. KCTD: A new gene family involved in neurodevelopmental and neuropsychiatric disorders. CNS Neurosci Ther. 2019;25:887-902. https://pubmed.ncbi.nlm.nih.gov/31197948/