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Hypoxia preconditioning elicit differential response in tissue-specific MSCs via immunomodulation and exosomal secretion

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Abstract

Mesenchymal stromal cells (MSCs) are emerging as an ideal candidate for regenerative medicine. It is known that the culture conditions impact the cellular properties of MSCs and their therapeutic behavior. Moreover, maintenance of MSCs in low oxygen tension for a short duration has shown to be beneficial for MSCs as it is similar to that of their physiological niche. However, the precise mechanism through which hypoxia pre-conditioning affects MSCs is not clear yet. Thus, in this study, we have investigated the effect of hypoxia exposure (1% O2) on tissue-specific MSCs over a period of time under serum-free culture conditions and evaluated the changes in expression of immuno-modulatory molecules and exosome biogenesis and secretion markers. It was observed that all MSCs responded differentially towards hypoxia exposure as indicated by the expression of HIF-1α. Moreover, this short-term exposure did not induce any changes in MSCs cellular morphology, proliferation rate, and surface marker profiling. In addition, we observed an enhancement in the expression of immunomodulatory factors (HLA-G, PGE-2, and IDO) after hypoxia exposure of 12 to 24 h in all tissue-specific MSCs. Interestingly, we have also observed the upregulation in exosome secretion that was further corelated to the upregulation of expression of exosome biogenesis and secretion markers (ALIX, TSG101, RAB27a, RAB27b). Though there was a differential response of MSCs where WJ-MSCs and BM-MSCs showed upregulation of these markers at 6–12 h of hypoxia pre-conditioning, while AD-MSCs showed similar changes beyond 24 h of hypoxia exposure.

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All data generated or analyzed during this study are included in this published article and its supplementary information files.

References

  • Adolfsson E, Helenius G, Friberg Ö, Samano N, Frøbert O, Johansson K (2020) Bone marrow-and adipose tissue-derived mesenchymal stem cells from donors with coronary artery disease; growth, yield, gene expression and the effect of oxygen concentration. Scand J Clin Lab Invest 80(4):318–326

    Article  CAS  Google Scholar 

  • Alhattab D, Jamali F, Ali D, Hammad H, Adwan S, Rahmeh R, Awidi A (2019) An insight into the whole transcriptome profile of four tissue-specific human mesenchymal stem cells. Regen Med 14(9):841–865

    Article  CAS  Google Scholar 

  • Amobi A, Qian F, Lugade AA, Odunsi K (2017) Tryptophan catabolism and cancer immunotherapy targeting IDO mediated immune suppression. Tumor Imm Microenviron Cancer Progress Cancer Ther 129–144

  • Antebi B, Rodriguez LA, Walker KP, Asher AM, Kamucheka RM, Alvarado L, Cancio LC (2018) Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells. Stem Cell Res Ther 9(1):1–15

    Article  Google Scholar 

  • Aryani A, Denecke B (2016) Exosomes as a nanodelivery system: a key to the future of neuromedicine?. Mol Neurobiol 53(2):818–834. https://doi.org/10.1007/s12035-014-9054-5

    Article  CAS  PubMed  Google Scholar 

  • Babst M (2011) MVB vesicle formation: ESCRT-dependent, ESCRT-independent and everything in between. Curr Opin Cell Biol 23(4):452–457. https://doi.org/10.1016/j.ceb.2011.04.008

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Burnham AJ, Foppiani EM, Horwitz EM (2020) Key metabolic pathways in MSC-mediated immunomodulation: implications for the prophylaxis and treatment of graft versus host disease. Front Immunol 11

  • Choi JR, Yong KW, Wan Safwani WK (2017) Effect of hypoxia on human adipose-derived mesenchymal stem cells and its potential clinical applications. Cell Mol Life Sci 74(14):2587–2600

    Article  CAS  Google Scholar 

  • Datta A, Kim H, McGee L, Johnson AE, Talwar S, Marugan J, Abdel-Mageed AB (2018) High-throughput screening identified selective inhibitors of exosome biogenesis and secretion: a drug repurposing strategy for advanced cancer. Sci Rep 8(1):1–13

    Article  CAS  Google Scholar 

  • Dionigi B, Ahmed A, Pennington EC, Zurakowski D, Fauza DO (2014) A comparative analysis of human mesenchymal stem cell response to hypoxia in vitro: Implications to translational strategies. J Pediatr Surg 49(6):915–918

    Article  Google Scholar 

  • Elahi FM, Farwell DG, Nolta JA, Anderson JD (2020) Preclinical translation of exosomes derived from mesenchymal stem/stromal cells. Stem Cells 38(1):15–21

    Article  Google Scholar 

  • Ferreira JR, Teixeira GQ, Santos SG, Barbosa MA, Almeida-Porada G, Gonçalves RM (2018) Mesenchymal stromal cell secretome: influencing therapeutic potential by cellular pre-conditioning. Front Immunol 9:2837. https://doi.org/10.3389/fimmu.2018.02837

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ferreira LM, Meissner TB, Tilburgs T, Strominger JL (2017) HLA-G: at the interface of maternal–fetal tolerance. Trends Immunol 38(4):272–286

    Article  CAS  Google Scholar 

  • Gebler A, Zabel O, Seliger B (2012) The immunomodulatory capacity of mesenchymal stem cells. Trends Mol Med 18(2):128–134

    Article  CAS  Google Scholar 

  • Gupta S, Rawat S, Arora V, Kottarath SK, Dinda AK, Vaishnav PK, Mohanty S (2018) An improvised one-step sucrose cushion ultracentrifugation method for exosome isolation from culture supernatants of mesenchymal stem cells. Stem Cell Res Ther 9(1):1–11

    Article  CAS  Google Scholar 

  • Gyöngyösi M, Hemetsberger R, Posa A, Charwat S, Pavo N, Petnehazy Ö, Maurer G (2010) Hypoxia-inducible factor 1-alpha release after intracoronary versus intramyocardial stem cell therapy in myocardial infarction. J Cardiovasc Transl Res 3(2):114–121

    Article  Google Scholar 

  • Holzwarth C, Vaegler M, Gieseke F, Pfister SM, Handgretinger R, Kerst G, Müller I (2010) Low physiologic oxygen tensions reduce proliferation and differentiation of human multipotent mesenchymal stromal cells. BMC Cell Biol 11(1):1–11

    Article  Google Scholar 

  • Kakkar A, Nandy SB, Gupta S, Bharagava B, Airan B, Mohanty S (2019) Adipose tissue derived mesenchymal stem cells are better respondents to TGFβ1 for in vitro generation of cardiomyocyte-like cells. Mol Cell Biochem 460(1):53–66

    Article  CAS  Google Scholar 

  • Kim DS, Ko YJ, Lee MW, Park HJ, Park YJ, Kim DI, Yoo KH (2016) Effect of low oxygen tension on the biological characteristics of human bone marrow mesenchymal stem cells. Cell Stress Chaperones 21(6):1089–1099

    Article  CAS  Google Scholar 

  • Kwon A, Kim Y, Kim M, Kim J, Choi H, Jekarl DW, Park IY (2016) Tissue-specific differentiation potency of mesenchymal stromal cells from perinatal tissues. Sci Rep 6(1):1–11

    Article  Google Scholar 

  • Kwon SY, Chun SY, Ha YS, Kim DH, Kim J, Song PH, Kwon TG (2017) Hypoxia enhances cell properties of human mesenchymal stem cells. Tissue Eng Regenerat Med 14(5):595–604

    Article  CAS  Google Scholar 

  • Larios J, Mercier V, Roux A, Gruenberg J (2020) ALIX-and ESCRT-III–dependent sorting of tetraspanins to exosomes. J Cell Biol 219(3)

  • Lee JH, Ha DH, Go HK, Youn J, Kim HK, Jin RC, Yi YW (2020) Reproducible large-scale isolation of exosomes from adipose tissue-derived mesenchymal stem/stromal cells and their application in acute kidney injury. Int J Mol Sci 21(13):4774

    Article  CAS  Google Scholar 

  • Lee JH, Yoon YM, Lee SH (2017) Hypoxic preconditioning promotes the bioactivities of mesenchymal stem cells via the HIF-1α-GRP78-Akt axis. Int J Mol Sci 18(6):1320

    Article  Google Scholar 

  • Li SP, Lin ZX, Jiang XY, Yu XY (2018) Exosomal cargo-loading and synthetic exosome-mimics as potential therapeutic tools. Acta Pharmacol Sin 39(4):542–551

    Article  CAS  Google Scholar 

  • Luan X, Sansanaphongpricha K, Myers I, Chen H, Yuan H, Sun D (2017) Engineering exosomes as refined biological nanoplatforms for drug delivery. Acta Pharmacol Sin 38(6):754–763

    Article  CAS  Google Scholar 

  • Pacary E, Legros H, Valable S, Duchatelle P, Lecocq M, Petit E, Bernaudin M (2006) Synergistic effects of CoCl2 and ROCK inhibition on mesenchymal stem cell differentiation into neuron-like cells. J Cell Sci 119(13):2667–2678

    Article  CAS  Google Scholar 

  • Paliwal S, Kakkar A, Sharma R, Airan B, Mohanty S (2017) Differential reduction of reactive oxygen species by human tissue-specific mesenchymal stem cells from different donors under oxidative stress. J Biosci 42(3):373–382

    Article  CAS  Google Scholar 

  • Panigrahi GK, Praharaj PP, Peak TC, Long J, Singh R, Rhim JS, Deep G (2018) Hypoxia-induced exosome secretion promotes survival of African-American and Caucasian prostate cancer cells. Sci Rep 8(1):1–13

    Google Scholar 

  • Rawat S, Srivastava P, Prabha P, Gupta S, Kanga U, Mohanty S (2018) A comparative study on immunomodulatory potential of tissue specific hMSCs: Role of HLA-G

  • Ren G, Su J, Zhang L, Zhao X, Ling W, L’huillie A, Shi Y (2009) Species variation in the mechanisms of mesenchymal stem cell-mediated immunosuppression. Stem Cells 27(8):1954–1962

    Article  CAS  Google Scholar 

  • Rezaie J, Nejati V, Khaksar M, Oryan A, Aghamohamadzadeh N, Shariatzadeh MA, Mehranjani MS (2018) Diabetic sera disrupted the normal exosome signaling pathway in human mesenchymal stem cells in vitro. Cell Tissue Res 374(3):555–565

    Article  CAS  Google Scholar 

  • Roemeling-van Rhijn M, Mensah FK, Korevaar SS, Leijs MJ, van Osch GJ, Ijzermans JN, Betjes MG, Baan CC, Weimar W, Hoogduijn MJ (2013) Effects of hypoxia on the immunomodulatory properties of adipose tissue-derived mesenchymal stem cells. Front Immunol 4:203. https://doi.org/10.3389/fimmu.2013.00203

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ruan XF, Ju CW, Shen Y, Liu YT, Kim IM, Yu H, Tang Y (2018) Suxiao Jiuxin pill promotes exosome secretion from mouse cardiac mesenchymal stem cells in vitro. Acta Pharmacol Sin 39(4):569–578

    Article  CAS  Google Scholar 

  • Saparov A, Ogay V, Nurgozhin T, Jumabay M, Chen WC (2016) Preconditioning of human mesenchymal stem cells to enhance their regulation of the immune response. Stem Cells Int 2016:3924858. https://doi.org/10.1155/2016/3924858

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sareen N, Abu-El-Rub E, Ammar HI, Yan W, Sequiera GL, ShamsEldeen AM, Dhingra S (2020) Hypoxia-induced downregulation of cyclooxygenase 2 leads to the loss of immunoprivilege of allogeneic mesenchymal stem cells. FASEB J 34(11):15236–15251

    Article  CAS  Google Scholar 

  • Schmidt SK, Ebel S, Keil E, Woite C, Ernst JF, Benzin AE, Rupp J, Däubener W (2013) Regulation of IDO activity by oxygen supply: inhibitory effects on antimicrobial and immunoregulatory functions. PLoS One 8(5):e63301. https://doi.org/10.1371/journal.pone.0063301

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shi X, Zhang G, Sun H, Bai Y, Liu Y, Zhang W, Xiao H (2018) Effects of over-expression of HIF-1alpha in bone marrow-derived mesenchymal stem cells on traumatic brain injury. Eng Life Sci 18(6):401–407

    Article  CAS  Google Scholar 

  • Singh M, Vaishnav PK, Dinda AK, Mohanty S (2020) Evaluation of priming efficiency of forskolin in tissue-specific human mesenchymal stem cells into dopaminergic neurons: an in vitro comparative study. Cells 9(9):2058

    Article  CAS  Google Scholar 

  • Teixeira FG, Panchalingam KM, Anjo SI, Manadas B, Pereira R, Sousa N, Behie LA (2015) Do hypoxia/normoxia culturing conditions change the neuroregulatory profile of Wharton Jelly mesenchymal stem cell secretome?. Stem Cell Res Ther 6(1):1–14

    Article  CAS  Google Scholar 

  • Tschuschke M, Kocherova I, Bryja A, Mozdziak P, Angelova Volponi A, Janowicz K, Sibiak R, Piotrowska-Kempisty H, Iżycki D, Bukowska D, Antosik P, Shibli JA, Dyszkiewicz-Konwińska M, Kempisty B (2020) Inclusion biogenesis, methods of isolation and clinical application of human cellular exosomes. J Clin Med 9(2):436. https://doi.org/10.3390/jcm9020436

    Article  CAS  PubMed Central  Google Scholar 

  • Wachowska M, Stachura J, Tonecka K, Fidyt K, Braniewska A, Sas Z, Muchowicz A (2020) Inhibition of IDO leads to IL-6-dependent systemic inflammation in mice when combined with photodynamic therapy. Cancer Immunol Immunother 69(6):1101–1112

    Article  CAS  Google Scholar 

  • Wang J, Bonacquisti EE, Brown AD, Nguyen J (2020) Boosting the biogenesis and secretion of mesenchymal stem cell-derived exosomes. Cells 9(3):660

    Article  CAS  Google Scholar 

  • Weidemann A, Johnson RS (2008) Biology of HIF-1 α. Cell Death Differ 15(4):621–627

    Article  CAS  Google Scholar 

  • Wobma HM, Kanai M, Ma SP, Shih Y, Li HW, Duran-Struuck R, Vunjak-Novakovic G (2018) Dual IFN-γ/hypoxia priming enhances immunosuppression of mesenchymal stromal cells through regulatory proteins and metabolic mechanisms. J Immunol Regenerat Med 1:45–56

    Article  Google Scholar 

  • Zhou X, Hong Y, Zhang H, Li X (2020) Mesenchymal stem cell senescence and rejuvenation: current status and challenges. Front Cell Developmen Biol 8

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Funding

The work has been supported by the All-India Institute of Medical Sciences, New Delhi, and the BIRAC-PACE (BT/AIR0987/PACE-19/19), Department of Biotechnology, Government of India and DST (EMR/2017/000229).

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  1. Suchi Gupta and Sonali Rawat have contributed equally to this manuscript.

Authors and Affiliations

  1. Stem Cell Facility, DBT-Centre of Excellence for Stem Cell Research), All India Institute of Medical Sciences, 1stFloor, ORBO Complex, Ansari Nagar, New Delhi, India

    Suchi Gupta, Sonali Rawat, Vishnu Krishnakumar, E. Pranshu Rao & Sujata Mohanty

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  1. Suchi Gupta
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  2. Sonali Rawat
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  3. Vishnu Krishnakumar
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  4. E. Pranshu Rao
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Contributions

Material preparation, data collection, and analysis were performed by SG and SR. Experiments were performed by SG, SR, VK, and EPR. The first draft of the manuscript was written by SG and all authors read and approved the final manuscript. SR contributed to analysis of data, paper writing, and first revision. SM contributed to supervision of experiments and critical revision of the manuscript.

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Correspondence to Sujata Mohanty.

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Mesenchymal stromal cells used in this study were isolated from healthy donors with written consent after obtaining ethical clearance (Ref No. ICSCR/34/15(R)) from Institutional Committee for Stem Cell research (ICSCR), AIIMS, New Delhi.

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Gupta, S., Rawat, S., Krishnakumar, V. et al. Hypoxia preconditioning elicit differential response in tissue-specific MSCs via immunomodulation and exosomal secretion. Cell Tissue Res 388, 535–548 (2022). https://doi.org/10.1007/s00441-022-03615-y

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