Increased transgene expression mediated by recombinant adeno-associated virus in human neuroglia cells under microgravity conditions

Feiyi Sun, Jiewen Zhang, Chengwei Ma, Yaxi Zhang, Yali Li, Lan Zhang, Yulin Deng, Hong Ma


The space environment has the special characteristics of radiation, noise particularity and weightlessness, all of which have adverse effects on astronauts’ muscles, bones, neurons and immune system. Some reports have shown that chemotherapy and radiotherapy can increase the activity of the recombinant adeno-associated virus (AAV) which is widely used in gene therapy. In this paper, recombinant AAV2 (rAAV2) was first packaged with the enhanced green fluorescence protein (eGFP) gene and used to infect neuroglia cells including the U87 and U251 cell lines, under microgravity conditions; it was then detected by fluorescence microscopy and flow cytometry. The results show that microgravity affects the adhesion ability of cells, promotes transgene expression induced by rAAV2 and causes changes of viral infection receptors at different time points. These findings broaden the current understanding of the microgravity effects on rAAV, with significant implications in gene therapy and the mechanisms of increased virus pathogenicity under space microgravity.



microgravity; Adeno-associated virus; adhesion ability; gene expression; pathogenicity


Aleshcheva G, Wehland M, Sahana J, Bauer J, Corydon TJ, Hemmersbach R, Frett T, Egli M, Infanger M, Grosse J & Grimm D 2015 Moderate alterations of the cytoskeleton in human chondrocytes after short-term microgravity produced by parabolic flight maneuvers could be prevented by up-regulation of BMP-2 and SOX-9. FASEB J 29 2303-2314

Atchison RW, Casto BC & Hammon WMD 1965 Adenovirus-associated defective virus particles. Science 149 754-756

Bartlett JS, Samulski RJ & McCown TJ. 1998 Selective and rapid uptake of adeno-associated virus type2 in brain. Human Gene Ther 9 1181-1186

Bishop JR, Schuksz M & Esko JD 2007 Heparan sulphate proteoglycans fine-tune mammalian physiology. Nature 446 1030-1037

Bonten EJ, Yogalingam G, Hu H, Gomero E, van de Vlekkert D & d'Azzo A 2013 Chaperone-mediated gene therapy with recombinant AAV-PPCA in a new mouse model of type I sialidosis. Biochim Biophys Acta 1832 1784-1792

Casto BC, Atchison RW & Hammon WM 1967 Studies on the relationship between adeno-associated virus type I (AAV- 1) and adenoviruses. I. Replication of AAV-1 in certain cell cultures and its effect on helper adenovirus. Virology 32 52-59

Chongzhen Wang, Hui Chen, Haiying Luo,Linnan Zhu, Yang Zhao, Hongling Tian, Ruoyu Wang, Peng Shang & Yong Zhao 2015 Microgravity activates p38 MAPK-C/EBPβ pathway to regulate the expression of arginase and inflammatory cytokines in macrophages. Inflammation Res 64 303-311

Chopra V, Fad AA, Sha J, Chopra S, Galindo CL & Chopra AK 2006 Alterations in the virulence potential of enteric pathogens and bacterial-host cell interactions under simulated microgravity conditions. J Toxicol Environ 69 1345-1370

Connor B, Sun Y, von Hieber D, Tang SK, Jones KS & Maucksch C 2016 AAV1/2-mediated BDNF gene therapy in a transgenic rat model of Huntington's disease. Gene Ther 23 283-295

Crucian BE, Stowe RP, Mehta SK, Yetman DL, Leal MJ, Quiriarte HD, Pierson DL & Sams CF 2009 Immune status, latent viral reactivation, and stress during long-duration head-down bed rest. Aviat Space Environ Med 80 37-44

Flotte T R 2004 Gene therapy progress and prospects: Recombinant adeno-associated virus (rAAV2) vectors. Gene Ther 11 805-810

Fuse A & Sato T 2004 Effect of microgravity changes on virus infection in mice. J Gravit Physiol 11 p65-66

Goldstein MA, Edwards RJ & Schroeter J P 1992 Cardiac morphology after conditions of microgravity during COSMOS 2044. J Appl Physiol 73 94-100

Gopalakrishnan R, Genc KO, Rice AJ, Lee SM, Evans HJ, Maender CC, Ilaslan H & Cavanagh PR 2010 Muscle volume, strength, endurance, and exercise loads during 6-month missions in space. Aviat Space Environ Med 81 91-102

Grosse J, Wehland M, Pietsch J, Ma X, Ulbrich C, Schulz H, Saar K, Hübner N, Hauslage J, Hemmersbach R, Braun M, van Loon J, Vagt N, Infanger M, Eilles C, Egli M, Richter P, Baltz T, Einspanier R, Sharbati S & Grimm D 2012 Short-term weightlessness produced by parabolic flight maneuvers altered gene expression patterns in human endothelial cells. FASEB J 26 639-655

Hoson T, Kamisaka S, Masuda Y & Yamashita M 1992 Changes in plant growth processes under microgravity condition simulated by a three-dimensional clinostat. Bot Mag 1077 53-70

Hudry E, Martin C, Gandhi S, György B, Scheffer DI, Mu D, Merkel SF, Mingozzi F, Fitzpatrick Z, Dimant H, Masek M, Ragan T, Tan S, Brisson AR, Ramirez SH, Hyman BT & Maguire CA 2016 Exosome-associated AAV vector as a robust and convenient neuroscience tool. Gene Ther 11 1-27

Hughes-Fulford M, Chang TT, Martinez EM & Li CF 2015 Spaceflight alters expression of microRNA during T-cell activation. FASEB J 29 4893-900

Humme S, Work LM, Janicki H, Hallek M, Baker AH & Büning H 2006 Heparan sulfate proteoglycan binding properties of adeno-associated virus retargeting mutants and consequences for their in vivo tropism. J Virol 80 7265-7269

Ingber D 1999 How cell (might) sense microgravity. FASEB J 13 3-15

Keyak JH, Koyama AK, LeBlanc A, Lu Y & Lang TF 2009 Reduction in proximal femoral strength due to long-duration spaceflight. Bone 44 449-453

King JA, Dubielzig R, Grimm D & Kleinschmidt JA 2001 DNA helicase-mediated packaging of adeno-associated virus type 2 genomes into preformed capsids. EMBO J 20 3282-3291

Kiss JZ 2014 Plant biology in reduced gravity on the Moon and Mars. Plant Biol 16 12-17

Loring HS, ElMallah MK & Flotte TR 2016 Development of RAAV2-CFTR: History of the First RAAV2 Vector Product to be used in Humans. Hum Gene Ther Methods 27 1-10

Lowrey CR, Perry SD, Strzalkowski ND, Williams DR, Wood SJ & Bent LR 2014 Selective skin sensitivity changes and sensory reweighting following short-duration space flight. J Appl Physiol 116 683-692

Luo J, Luo Y, Sun J, Zhou Y, Zhang Y & Yang X 2015 Adeno-associated virus-mediated cancer gene therapy: Current status. Cancer Let 356 347-356

Ma H, Liu Y, Liu S, Xu R & Zheng D 2005 Oral adeno-associated virus-sTRAIL gene therapy suppresses human hepatocellular carcinoma growth in mice. Hepatology 42 1355-1363

Madry H & Cucchiarini M 2016 Gene therapy for human osteoarthritis: principles and clinical translation. Expert Opin Biol Ther 16 331-346

Manno CS, Chew AJ, Hutchison S, Larson PJ, Herzog RW, Arruda VR, Tai SJ, Ragni MV, Thompson A, Ozelo M, Couto LB, Leonard DG, Johnson FA, McClelland A, Scallan C, Skarsgard E, Flake AW, Kay MA, High KA & Glader B 2003 AAV-mediated factor IX gene transfer to skeletal muscle in patients with severe hemophilia B. Blood 101 2963-2972

Manno CS, Pierce GF, Arruda VR, Glader B, Ragni M, Rasko JJ, Ozelo MC, Hoots K, Blatt P, Konkle B, Dake M, Kaye R, Razavi M, Zajko A, Zehnder J, Rustagi PK, Nakai H, Chew A, Leonard D, Wright JF, Lessard RR, Sommer JM, Tigges M, Sabatino D, Luk A, Jiang H, Mingozzi F, Couto L, Ertl HC, High KA & Kay MA 2006 Successful transduction of liver in hemophilia by AAV-Factor IX and limitations imposed by the host immune response. Nat Med 12 342-347

Meyers C, Alam S, Mane M & Hermonat PL 2001 Altered biology of adeno-associated virus type 2 and human papillomavirus during dual infection of natural host tissue. Virology 287 30-39

Morgan JL, Zwart SR, Heer M, Ploutz-Snyder R, Ericson K & Smith SM 2012 Bone metabolism and nutritional status during 30-day head-down-tilt bed rest. J Appl Physiol 113 1519-1529

Nickerson CA, Ott CM, Mister SJ, Morrow BJ, Burns-Keliher L & Pierson DL 2000 Microgravity as a novel environmental signal affecting salmonella enterica serovar typhimurium virulence. Infect Immun 68 3147-3152

Perabo L, Goldnau D, White K, Endell J, Boucas J & Russell W C 2009 Adeno viruses: update on structure and function. J Gen Virol 90 1-20

Philpott DE, Kato K & Miquel J 1992 Ultrastructural and cellular mechanisms in myocardial deconditioning in weightlessness. Adv Space Biol Med 2 83-112

Platts SH, Ziegler MG, Waters WW & Meck JV 2006 Hemodynamic effects of midodrine after spaceflight in astronauts without orthostatic hypotension. Aviat Space Eviron Med 77 429-433

Qing K, Mah C, Hansen J, Zhou S, Dwarki V & Srivastava A 1999 Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2. Nat Med 5 71-77

Salva MZ, Himeda CL, Tai PW, Nishiuchi E, Gregorevic P, Allen JM, Finn EE, Nguyen QG, Blankinship MJ, Meuse L, Chamberlain JS & Hauschka SD 2007 Design of tissue-specific regulatory cassettes for high-level rAAV2-mediated expression in skeletal and cardiac muscle. Mol Ther 15 320-329

Santiago-Ortiz JL & Schaffer DV 2016 Adeno-associated virus (AAV) vectors in cancer gene therapy. J Control Release 16 1-12

Zwart SR, Pierson D, Mehta S, Gonda S & Smith SM 2010 Capacity of omega-3 fatty acids or eicosapentaenoic acid to counteract weightlessness-induced bone loss by inhibiting NF-kappaB activation: from cells to bed rest to astronauts. J Bone Miner Res 25 1049-1057

Sato A, Kumei Y, Sato K, Hongo T, Hamazaki T, Masuda I, Nakajima T, Ohmura T, Kaiho M, Sato T & Wake K 2001 Studies on the effects of microgravity on the ultra-structure and function of cultured mammalian cells. Biol Sci Space 15 61-63

Schlehofer JR, Ehrbar M & zur Hausen H 1986 Vaccinia, herpes simpex, and carcinogen induced DNA amplification in a human cell line and support replication of a helpervirus dependent parvovirus. Virology 152 110-117

Schmidt M, Voutetakis A, Afione S, Zheng C, Mandikian D & Chiorini JA. 2008 Adeno-associated virus type12(AAV12):a novel AAV serotype with sialic acid and heparan sulfate proteoglycan-indepandent transduction activity. J Virol 82 1399-1406

Seisenberger G, Ried MU, Endress T, Büning H, Hallek M & Bräuchle C 2001 Real-time single-molecule imaging of the infection pathway of an adeno-associated virus. Science 294 1929-1932

Smith R 2008 Adeno-associated virus integration: virus versus vector. Gene Ther 15 817-822

Smith SM, Abrams SA, Davis-Street JE, Heer M, O'Brien KO, Wastney ME & Zwart SR 2014 Fifty years of human space travel: implications for bone and calcium research. Annu Rev Nutr 34 377-400

Sonnenfeld G & Shearer WT 2002 Immune function during space flight. Nutrition 18 899-903

Appel SH, Beers DR & Henkel JS 2010 T cell-microglial dialogue in Parkinson’s disease and amyotrophic lateral sclerosis: are we listening?. Trends Immunol 31 7-17

Stenger MB, Brown AK, Lee SM, Locke JP & Platts SH 2010 Gradient compression garments as a countermeasure to post-spaceflight orthostatic intolerance. Aviat Space Environ Med 81 883-887

Stowe RP, Kozlova EV, Sams CF, Pierson DL & Walling DM 2011 Latent and lytic Epstein-Barr virus gene expression in the peripheral blood of astronauts. J Med Virol 83 1071-1077

Stowe RP, Pierson DL & Barrett AD 2001 Elevated stress hormone levels relate to Epstein-Barr virus reactivation in astronauts psychosom. Med 63 891-895

Summerford C, Bartlett JS & Samulski RJ 1998 Membrane-associated heparin sulfate proteoglycan is a receptor for adeno-associated virus type 2 virions. J Virol 72 1438-1445

Summerford C, Bartlett JS & Samulski RJ 1999 AlphaVbeta5 integrin: a co-receptor for adeno-associated virus type 2 infection. Nat Med 5 78-82

Tesch PA, Berg HE, Bring D, Evans HJ, LeBlanc AD 2005 Effects of 17-day spaceflight on knee extensor muscle function and size. Eur J Appl Physiol 93 463-468

Wu Z, Asokan A & Samulski RJ 2006 Adeno-associated virus serotypes:vector toolkit for human gene therapy. Mol Ther 14 316-327

Zhou X, Shen L, Liu L, Wang C, Qi W, Zhao A, Wu X & Li B 2016 Preclinical safety evaluation of recombinant adeno-associated virus 2 vector encoding human tumor necrosis factor receptor-immunoglobulin Fc fusion gene. Hum Vaccin Immunother 2 1-24

Full Text: PDF


  • There are currently no refbacks.

Copyright © 2017 Lorem Ipsum Press