Abstract
We recently showed that membrane progesterone receptor α (mPRα/PAQR7) promotes pro-regenerative effects in Schwann cell-like adipose stem cells (SCL-ASC), an alternative model to Schwann cells for the promotion of peripheral nerve regeneration. In this study, we investigated how mPRα activation with the mPR-specific agonist Org OD 02–0 in SCL-ASC affected regenerative parameters in two neuronal cell lines, IMR-32 and SH-SY-5Y. In a series of conditioned medium experiments, we found that mPR activation of SCL-ASC led to increased neurite outgrowth, protection from cell death and increased expression of peripheral nerve regeneration markers (CREB3, ATF3, GAP43) in neuronal cell lines. These effects were stronger than the ones observed with the conditioned medium from untreated SCL-ASC. The addition of Org OD 02–0 to the untreated cell medium mimicked the effects of mPR activation of SCL-ASC on cell death, but not on neurite outgrowth. Therefore, the effect of Org OD 02–0 on neurite outgrowth is SCL-ASC-dependent, while its effect on cell survivability is likely due to the direct activation of mPRs on neuronal cells. SCL-ASC transfection with mPRα siRNA showed that this isoform is responsible for the beneficial effect on neurite outgrowth. Further experiments showed that SCL-ASC-dependent outcomes likely involved the release of BDNF and IGF-2 from these cells. The beneficial mPRα effect on neurite outgrowth was confirmed in co-culture conditions. These findings strengthen the hypothesis that mPRα could play a pro-regenerative role in SCL-ASC and be a therapeutic target for the promotion of peripheral nerve regeneration.
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References
Castelnovo LF, Bonalume V, Melfi S et al (2017) Schwann cell development, maturation and regeneration: a focus on classic and emerging intracellular signaling pathways. Neural Regen Res 12:1013–1023. https://doi.org/10.4103/1673-5374.211172
Castelnovo LF, Caffino L, Bonalume V et al (2020) Membrane progesterone receptors (mPRs/PAQRs) differently regulate migration, proliferation, and differentiation in rat Schwann cells. J Mol Neurosci 70:433–448. https://doi.org/10.1007/s12031-019-01433-6
Castelnovo LF, Magnaghi V, Thomas P (2019) Expression of membrane progesterone receptors (mPRs) in rat peripheral glial cell membranes and their potential role in the modulation of cell migration and protein expression. Steroids 142:6–13. https://doi.org/10.1016/j.steroids.2017.09.009
Castelnovo LF, Thomas P (2021) Membrane progesterone receptor α (mPRα/PAQR7) promotes migration, proliferation and BDNF release in human Schwann cell-like differentiated adipose stem cells. Mol Cell Endocrinol 531:111298. https://doi.org/10.1016/j.mce.2021.111298
Castelnovo LF, Thomas P, Magnaghi V (2021) Membrane progesterone receptors (mPRs/PAQRs) in Schwann cells represent a promising target for the promotion of neuroregeneration. Neural Regen Res 16:281–282. https://doi.org/10.4103/1673-5374.290885
Chandran V, Coppola G, Nawabi H et al (2016) A systems-level analysis of the peripheral nerve intrinsic axonal growth program. Neuron 89:956–970. https://doi.org/10.1016/j.neuron.2016.01.034
Chen Y, Shen J, Ma C et al (2020) Skin-derived precursor Schwann cells protect SH-SY5Y cells against 6-OHDA-induced neurotoxicity by PI3K/AKT/Bcl-2 pathway. Brain Res Bull 161:84–93. https://doi.org/10.1016/j.brainresbull.2020.03.020
Ching RC, Wiberg M, Kingham PJ (2018) Schwann cell-like differentiated adipose stem cells promote neurite outgrowth via secreted exosomes and RNA transfer. Stem Cell Res Ther 9:266. https://doi.org/10.1186/s13287-018-1017-8
de Luca AC, Faroni A, Reid AJ (2015) Dorsal root ganglia neurons and differentiated adipose-derived stem cells: an in vitro co-culture model to study peripheral nerve regeneration. J Vis Exp 96:52543. https://doi.org/10.3791/52543
Dezawa M, Takahashi I, Esaki M et al (2001) Sciatic nerve regeneration in rats induced by transplantation of in vitro differentiated bone-marrow stromal cells. Eur J Neurosci 14:1771–1776. https://doi.org/10.1046/j.0953-816x.2001.01814.x
Dressing GE, Alyea R, Pang Y, Thomas P (2012) Membrane progesterone receptors (mPRs) mediate progestin induced antimorbidity in breast cancer cells and are expressed in human breast tumors. Horm Cancer 3:101–112. https://doi.org/10.1007/s12672-012-0106-x
Endo T, Kadoya K, Kawamura D, Iwasaki N (2019) Evidence for cell-contact factor involvement in neurite outgrowth of dorsal root ganglion neurons stimulated by Schwann cells. Exp Physiol 104:1447–1454. https://doi.org/10.1113/EP087634
Faroni A, Mobasseri SA, Kingham PJ, Reid AJ (2015) Peripheral nerve regeneration: experimental strategies and future perspectives. Adv Drug Deliv Rev 82–83:160–167. https://doi.org/10.1016/j.addr.2014.11.010
Faroni A, Smith RJ, Reid AJ (2014) Adipose derived stem cells and nerve regeneration. Neural Regen Res 9:1341–1346. https://doi.org/10.4103/1673-5374.137585
Faroni A, Smith RJP, Lu L, Reid AJ (2016) Human Schwann-like cells derived from adipose-derived mesenchymal stem cells rapidly de-differentiate in the absence of stimulating medium. Eur J Neurosci 43:417–430. https://doi.org/10.1111/ejn.13055
Faroni A, Terenghi G, Magnaghi V (2012) Expression of functional γ-aminobutyric acid type A receptors in Schwann-like adult stem cells. J Mol Neurosci 47:619–630. https://doi.org/10.1007/s12031-011-9698-9
Jessen KR, Arthur-Farraj P (2019) Repair Schwann cell update: adaptive reprogramming, EMT, and stemness in regenerating nerves. Glia 67:421–437. https://doi.org/10.1002/glia.23532
Jessen KR, Mirsky R (2019) The success and failure of the Schwann cell response to nerve injury. Front Cell Neurosci 13:33. https://doi.org/10.3389/fncel.2019.00033
Jessen KR, Mirsky R, Lloyd AC (2015) Schwann cells: development and role in nerve repair. Cold Spring Harb Perspect Biol 7:a020487. https://doi.org/10.1101/cshperspect.a020487
Karteris E, Zervou S, Pang Y et al (2006) Progesterone signaling in human myometrium through two novel membrane G protein-coupled receptors: potential role in functional progesterone withdrawal at term. Mol Endocrinol 20:1519–1534. https://doi.org/10.1210/me.2005-0243
Kasubuchi M, Watanabe K, Hirano K et al (2017) Membrane progesterone receptor beta (mPRβ/Paqr8) promotes progesterone-dependent neurite outgrowth in PC12 neuronal cells via non-G protein-coupled receptor (GPCR) signaling. Sci Rep 7:5168. https://doi.org/10.1038/s41598-017-05423-9
Kelder J, Azevedo R, Pang Y et al (2010) Comparison between steroid binding to membrane progesterone receptor α (mPRα) and to nuclear progesterone receptor: correlation with physicochemical properties assessed by comparative molecular field analysis and identification of mPRα-specific agonists. Steroids 75:314–322. https://doi.org/10.1016/j.steroids.2010.01.010
Kim T-H, Yoon S-J, Lee W-C et al (2011) Protective effect of GCSB-5, an herbal preparation, against peripheral nerve injury in rats. J Ethnopharmacol 136:297–304. https://doi.org/10.1016/j.jep.2011.04.037
Kingham PJ, Kalbermatten DF, Mahay D et al (2007) Adipose-derived stem cells differentiate into a Schwann cell phenotype and promote neurite outgrowth in vitro. Exp Neurol 207:267–274. https://doi.org/10.1016/j.expneurol.2007.06.029
Kingham PJ, Kolar MK, Novikova LN et al (2014) Stimulating the neurotrophic and angiogenic properties of human adipose-derived stem cells enhances nerve repair. Stem Cells Dev 23:741–754. https://doi.org/10.1089/scd.2013.0396
Lopes CDF, Gonçalves NP, Gomes CP et al (2017) BDNF gene delivery mediated by neuron-targeted nanoparticles is neuroprotective in peripheral nerve injury. Biomaterials 121:83–96. https://doi.org/10.1016/j.biomaterials.2016.12.025
Matsuse D, Kitada M, Kohama M et al (2010) Human umbilical cord-derived mesenchymal stromal cells differentiate into functional Schwann cells that sustain peripheral nerve regeneration. J Neuropathol Exp Neurol 69:973–985. https://doi.org/10.1097/NEN.0b013e3181eff6dc
McGregor CE, English AW (2018) The role of BDNF in peripheral nerve regeneration: activity-dependent treatments and Val66Met. Front Cell Neurosci 12:522. https://doi.org/10.3389/fncel.2018.00522
Neill D, Hughes D, Edwardson JA et al (1994) Human IMR-32 neuroblastoma cells as a model cell line in Alzheimer’s disease research. J Neurosci Res 39:482–493. https://doi.org/10.1002/jnr.490390415
Pang Y, Dong J, Thomas P (2013) Characterization, neurosteroid binding and brain distribution of human membrane progesterone receptors δ and ε (mPRδ and mPRε) and mPRδ involvement in neurosteroid inhibition of apoptosis. Endocrinology 154:283–295. https://doi.org/10.1210/en.2012-1772
Parakala ML, Zhang Y, Modgil A et al (2019) Metabotropic, but not allosteric, effects of neurosteroids on GABAergic inhibition depend on the phosphorylation of GABAA receptors. J Biol Chem 294:12220–12230. https://doi.org/10.1074/jbc.RA119.008875
Peng J, Wang Y, Zhang L et al (2011) Human umbilical cord Wharton’s jelly-derived mesenchymal stem cells differentiate into a Schwann-cell phenotype and promote neurite outgrowth in vitro. Brain Res Bull 84:235–243. https://doi.org/10.1016/j.brainresbull.2010.12.013
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45. https://doi.org/10.1093/nar/29.9.e45
Piovesana R, Faroni A, Taggi M et al (2020) Muscarinic receptors modulate nerve growth factor production in rat Schwann-like adipose-derived stem cells and in Schwann cells. Sci Rep 10:7159. https://doi.org/10.1038/s41598-020-63645-w
Piovesana R, Faroni A, Tata A, Reid A (2021) Schwann-like adipose-derived stem cells as a promising therapeutic tool for peripheral nerve regeneration: effects of cholinergic stimulation. Neural Regen Res 16:1218. https://doi.org/10.4103/1673-5374.300433
Poplawski G, Ishikawa T, Brifault C et al (2018) Schwann cells regulate sensory neuron gene expression before and after peripheral nerve injury. Glia 66:1577–1590. https://doi.org/10.1002/glia.23325
Porzionato A, Barbon S, Stocco E et al (2019) Development of oxidized polyvinyl alcohol-based nerve conduits coupled with the ciliary neurotrophic factor. Materials (basel) 12:1996. https://doi.org/10.3390/ma12121996
Romeo-Guitart D, Leiva-Rodriguez T, Forés J, Casas C (2019) Improved motor nerve regeneration by SIRT1/Hif1a-mediated autophagy. Cells 8:1354. https://doi.org/10.3390/cells8111354
Sowa Y, Kishida T, Imura T et al (2016) Adipose-derived stem cells promote peripheral nerve regeneration in vivo without differentiation into Schwann-like lineage. Plast Reconstr Surg 137:318e–330e. https://doi.org/10.1097/01.prs.0000475762.86580.36
Stocco E, Barbon S, Lamanna A et al (2021) Bioactivated oxidized polyvinyl alcohol towards next-generation nerve conduits development. Polymers (basel) 13:3372. https://doi.org/10.3390/polym13193372
Thomas P, Pang Y (2012) Membrane progesterone receptors: evidence for neuroprotective, neurosteroid signaling and neuroendocrine functions in neuronal cells. Neuroendocrinology 96:162–171. https://doi.org/10.1159/000339822
Thomas P, Pang Y (2020) Anti-apoptotic actions of Allopregnanolone and Ganaxolone mediated through membrane progesterone receptors (PAQRs) in neuronal cells. Front Endocrinol (lausanne) 11:417. https://doi.org/10.3389/fendo.2020.00417
Thomas P, Pang Y, Dong J et al (2007) Steroid and G protein binding characteristics of the seatrout and human progestin membrane receptor alpha subtypes and their evolutionary origins. Endocrinology 148:705–718. https://doi.org/10.1210/en.2006-0974
Tohill M, Mantovani C, Wiberg M, Terenghi G (2004) Rat bone marrow mesenchymal stem cells express glial markers and stimulate nerve regeneration. Neurosci Lett 362:200–203. https://doi.org/10.1016/j.neulet.2004.03.077
Tulchinsky D, Hobel CJ, Yeager E, Marshall JR (1972) Plasma estrone, estradiol, estriol, progesterone, and 17-hydroxyprogesterone in human pregnancy. I. Normal Pregnancy Am J Obstet Gynecol 112:1095–1100. https://doi.org/10.1016/0002-9378(72)90185-8
Watanabe Y, Sasaki R, Matsumine H et al (2017) Undifferentiated and differentiated adipose-derived stem cells improve nerve regeneration in a rat model of facial nerve defect. J Tissue Eng Regen Med 11:362–374. https://doi.org/10.1002/term.1919
Welleford AS, Quintero JE, El SN et al (2020) RNA sequencing of human peripheral nerve in response to injury: distinctive analysis of the nerve repair pathways. Cell Transplant 29:963689720926157. https://doi.org/10.1177/0963689720926157
Xicoy H, Wieringa B, Martens GJM (2017) The SH-SY5Y cell line in Parkinson’s disease research: a systematic review. Mol Neurodegener 12:10. https://doi.org/10.1186/s13024-017-0149-0
Ye J, Coulouris G, Zaretskaya I et al (2012) Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics 13:134. https://doi.org/10.1186/1471-2105-13-134
Ying Z, Zhai R, McLean NA et al (2015) The unfolded protein response and cholesterol biosynthesis link Luman/CREB3 to regenerative axon growth in sensory neurons. J Neurosci 35:14557–14570. https://doi.org/10.1523/JNEUROSCI.0012-15.2015
Zhu Y, Bond J, Thomas P (2003a) Identification, classification, and partial characterization of genes in humans and other vertebrates homologous to a fish membrane progestin receptor. Proc Natl Acad Sci U S A 100:2237–2242. https://doi.org/10.1073/pnas.0436133100
Zhu Y, Rice CD, Pang Y et al (2003b) Cloning, expression, and characterization of a membrane progestin receptor and evidence it is an intermediary in meiotic maturation of fish oocytes. Proc Natl Acad Sci U S A 100:2231–2236. https://doi.org/10.1073/pnas.0336132100
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This research project was supported by The Morris L. Lichtenstein, Jr. Medical Research Foundation.
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Both authors contributed to the study’s conception and design. L.F.C. performed the experiments, wrote the original draft, and prepared the figures. P.T. obtained funding, supervised the project, and revised the manuscript. Both authors approved the final version of the manuscript.
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Castelnovo, L.F., Thomas, P. Membrane Progesterone Receptor α (mPRα/PAQR7) Promotes Survival and Neurite Outgrowth of Human Neuronal Cells by a Direct Action and Through Schwann Cell-like Stem Cells. J Mol Neurosci 72, 2067–2080 (2022). https://doi.org/10.1007/s12031-022-02057-z
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DOI: https://doi.org/10.1007/s12031-022-02057-z