{"id":7976,"date":"2022-05-03T10:47:39","date_gmt":"2022-05-03T14:47:39","guid":{"rendered":"https:\/\/mrm.research.mcgill.ca\/2022\/05\/mrm-insights-ipsc-derived-oligodendrocytes-a-model-for-regenerative-potential-in-multiple-sclerosis\/"},"modified":"2022-05-04T10:32:08","modified_gmt":"2022-05-04T14:32:08","slug":"mrm-insights-oligodendrocytes-derives-dipsc-un-modele-de-potentiel-de-regeneration-dans-la-sep","status":"publish","type":"post","link":"https:\/\/mrm.research.mcgill.ca\/fr\/2022\/05\/mrm-insights-oligodendrocytes-derives-dipsc-un-modele-de-potentiel-de-regeneration-dans-la-sep\/","title":{"rendered":"MRM Insights : Oligodendrocytes d\u00e9riv\u00e9s d\u2019IPSC \u2013 un mod\u00e8le de potentiel de r\u00e9g\u00e9n\u00e9ration dans la SEP ?"},"content":{"rendered":"
\"\"

Valerio Piscopo<\/p><\/div>\n

\"\"

Thomas Durcan<\/p><\/div>\n

\"\"

Gabriela Blaszczyk<\/p><\/div>\n

\"\"

Jack Antel<\/p><\/div>\n

Chaque mois, dans les MRM Insights<\/a>, un membre du R\u00e9seau MRM \u00e9crit sur les cellules souches et la m\u00e9decine r\u00e9g\u00e9n\u00e9rative d\u2019un point de vue diff\u00e9rent. Ce mois-ci, Jack Antel<\/strong>, Professeur de neurologie et de neurochirurgie au Neuro et \u00e0 l’Universit\u00e9 McGill, Gabriela Blaszczyk<\/strong>, \u00e9tudiante de 1er<\/sup> cycle dans son laboratoire, Thomas Durcan, <\/strong>Professeur agr\u00e9g\u00e9 au Neuro et \u00e0 l’Universit\u00e9 McGill, Directeur de la PDMPP (EDDU), et Valerio Piscopo<\/strong>, Associ\u00e9 de recherche \u00e0 la PDMPP, nous parlent des oligodendrocytes d\u00e9riv\u00e9s d’iPSC comme d’un mod\u00e8le du potentiel de r\u00e9g\u00e9n\u00e9ration dans la scl\u00e9rose en plaque<\/strong>.<\/p>\n

Oligodendrocytes d\u00e9riv\u00e9s d\u2019IPSC \u2013 un mod\u00e8le de potentiel de r\u00e9g\u00e9n\u00e9ration dans la SEP ?<\/h2>\n

Les oligodendrocytes (OL) sont une population de cellules gliales qui produisent de la my\u00e9line autour des axones, permettant une transduction efficace du signal dans le syst\u00e8me nerveux central (SNC). Les OL fournissent un soutien trophique essentiel aux neurones. La caract\u00e9ristique pathologique de la scl\u00e9rose en plaques (SEP) est la destruction de la my\u00e9line. La phase initiale de l\u2019\u00e9volution de la SEP est caract\u00e9ris\u00e9e par des rechutes r\u00e9currentes de dysfonctionnement neurologique r\u00e9sultant de l\u00e9sions d\u00e9my\u00e9linisantes associ\u00e9es \u00e0 une inflammation focale avec une \u00e9pargne relative des corps cellulaires des OL (1). La r\u00e9cup\u00e9ration apr\u00e8s ces rechutes est li\u00e9e au moins en partie \u00e0 la remy\u00e9linisation dans de telles l\u00e9sions. Au fil du temps, la maladie peut \u00e9voluer vers une phase progressive avec une aggravation progressive des sympt\u00f4mes neurologiques; cette phase est associ\u00e9e \u00e0 la perte de corps cellulaires des OL, \u00e0 l\u2019incapacit\u00e9 de r\u00e9g\u00e9n\u00e9rer la gaine de my\u00e9line et \u00e0 la perte neuronale subs\u00e9quente (1). La remy\u00e9linisation dans le SNC est g\u00e9n\u00e9ralement attribu\u00e9e au recrutement et \u00e0 la diff\u00e9renciation des cellules prog\u00e9nitrices oligodendrocytes (OPC) (2), bien que des OL pr\u00e9c\u00e9demment my\u00e9linisants puissent \u00e9galement y contribuer.<\/p>\n

Les m\u00e9canismes pr\u00e9cis qui sous-tendent les processus de blessure et de r\u00e9paration dans la SEP restent \u00e0 d\u00e9finir. Les constituants de l\u2019inflammation qui peuvent \u00eatre d\u00e9riv\u00e9s de l\u2019infiltration des cellules immunitaires, ainsi que des astrocytes activ\u00e9s et la microglie sont impliqu\u00e9s en tant que m\u00e9diateurs dans les processus de blessure (Fig 1). Les conditions dans le microenvironnement des l\u00e9sions de SEP cr\u00e9ent un stress m\u00e9tabolique sur les OL. Le succ\u00e8s de la r\u00e9paration de la my\u00e9line a \u00e9t\u00e9 li\u00e9 \u00e0 la production de facteurs trophiques essentiels et \u00e0 la r\u00e9solution des conditions inhibitrices locales. Des mod\u00e8les animaux, tels que l\u2019enc\u00e9phalomy\u00e9lite auto-immune exp\u00e9rimentale (EAE) et la d\u00e9my\u00e9linisation induite par les toxines, ont \u00e9t\u00e9 d\u00e9velopp\u00e9s pour \u00e9valuer \u00e0 la fois les processus de blessure et de r\u00e9paration associ\u00e9s \u00e0 la SEP.<\/p>\n

\"\"

Figure 1.<\/strong>Lib\u00e9ration de mol\u00e9cules pro-inflammatoires qui agissent sur les OL lors de la formation de l\u00e9sions de SEP. Fabriqu\u00e9 avec BioRender. Adapt\u00e9 de : Wu GF, Alvarez E. Neurol Clin. 2011. Levite M. J Neural Transm (Vienna). 2017. Frigo M, et al. Curr Med Chem. 2012<\/p><\/div>\n

Un d\u00e9fi permanent consiste \u00e0 d\u00e9montrer les caract\u00e9ristiques mol\u00e9culaires sp\u00e9cifiques des OL humains qui sous-tendent leur susceptibilit\u00e9 aux blessures et leur capacit\u00e9 \u00e0 effectuer des r\u00e9parations. L\u2019analyse directe des tissus touch\u00e9s par la SEP fournit des indices, mais elle est limit\u00e9e en ce qui concerne la d\u00e9finition des aspects dynamiques des processus de blessure et de r\u00e9paration. L\u2019acc\u00e8s aux OL humains primaires pour les \u00e9tudes fonctionnelles et mol\u00e9culaires est \u00e9galement limit\u00e9, bien qu\u2019il soit men\u00e9 dans le cadre de notre programme gr\u00e2ce aux efforts de collaboration des D\u00e9partements de neurochirurgie adulte et p\u00e9diatrique de McGill et du Programme de neuroimmunologie du Neuro (3).<\/p>\n

L\u2019une des approches \u00e9mergentes pour faire progresser notre compr\u00e9hension des propri\u00e9t\u00e9s de blessure \/ r\u00e9paration des OL humains implique l\u2019utilisation de cellules souches pluripotentes induites humaines (iPSC), car elles peuvent \u00eatre facilement d\u00e9riv\u00e9es de cellules somatiques de patients et peuvent se diff\u00e9rencier en tous les types de cellules (4). Cette approche est utilis\u00e9e par la collaboration du Programme de neuroimmunologie du Neuro, du programme de science ouverte du Neuro par l\u2019entremise du R\u00e9f\u00e9rentiel C-BIG (https:\/\/www.mcgill.ca\/neuro\/fr\/science-ouverte\/referentiel-c-big) et de l\u2019Unit\u00e9 de d\u00e9veloppement pr\u00e9coce de m\u00e9dicaments (PDMPP) avec le soutien du R\u00e9seau MRM. En utilisant un protocole pr\u00e9alablement \u00e9tabli (5), nous pouvons g\u00e9n\u00e9rer efficacement des OPC, ce qui nous permet d\u2019\u00e9tudier leur susceptibilit\u00e9 aux blessures et les m\u00e9canismes potentiels contribuant \u00e0 leur carence en r\u00e9paration dans des conditions telles que celles observ\u00e9es dans la SEP.<\/p>\n

Nous avons utilis\u00e9 les OPC d\u00e9riv\u00e9s d\u2019iPSC pour faire progresser les \u00e9tudes visant \u00e0 d\u00e9terminer si et comment les mol\u00e9cules pro-inflammatoires contribuent \u00e0 la disparition des OL et de leurs pr\u00e9curseurs. Parmi ces mol\u00e9cules, comme illustr\u00e9 \u00e0 la figure 1, on trouve le glutamate qui peut \u00eatre lib\u00e9r\u00e9 par des cellules Th17 activ\u00e9es ou des microglies r\u00e9sidentes; l\u2019interf\u00e9ron gamma (IFN\u03b3), qui peut \u00e9galement \u00eatre lib\u00e9r\u00e9 par les lymphocytes T activ\u00e9s, et le facteur de n\u00e9crose tumorale-alpha (TNF\u03b1) produit par les macrophages activ\u00e9s, les lymphocytes T et les cellules tueuses naturelles (NK) (6 – 8). Jusqu\u2019\u00e0 pr\u00e9sent, les effets de ces mol\u00e9cules ont \u00e9t\u00e9 explor\u00e9s in vivo dans des mod\u00e8les exp\u00e9rimentaux, in vitro en utilisant des lign\u00e9es d\u2019OL non humains, et dans nos propres \u00e9tudes in vitro utilisant des OL humains matures (9). Ces derni\u00e8res \u00e9tudes indiquent que les m\u00e9diateurs induisent la r\u00e9traction des neurites mais pas la mort cellulaire dans les cellules humaines, contrairement aux observations faites avec des cellules prog\u00e9nitrices non humaines. Les \u00e9tudes iPSC nous permettent d\u2019\u00e9valuer les effets de ces m\u00e9diateurs tout au long du spectre de la diff\u00e9renciation des cellules OL humaines. En utilisant cette approche, nous observons que l\u2019IFN\u03b3 et le TNF\u03b1 individuellement arr\u00eatent la diff\u00e9renciation des OPC vers les oligodendrocytes, plut\u00f4t que d\u2019initier des programmes de mort cellulaire, une d\u00e9couverte en accord avec Staros et coll. qui ont utilis\u00e9 une approche de reprogrammation g\u00e9n\u00e9tique pour g\u00e9n\u00e9rer des OL d\u00e9riv\u00e9s d\u2019iPSC (10). Ce syst\u00e8me de dosage permet d\u2019\u00e9valuer un large \u00e9ventail de mol\u00e9cules d\u00e9riv\u00e9es des cellules immunitaires (y compris les cellules B) (11) et des cellules gliales.<\/p>\n

Le suivi du stade de d\u00e9veloppement des OPC in vitro n\u00e9cessite des analyses immunocytochimiques complexes intermittentes. Pour nous permettre de suivre ces cellules de mani\u00e8re dynamique au fil du temps, le R\u00e9seau MRM a sp\u00e9cifiquement soutenu nos efforts pour g\u00e9n\u00e9rer une lign\u00e9e cellulaire iPSC exprimant un g\u00e8ne rapporteur de lign\u00e9e OL (SOX10), nous permettant de suivre la diff\u00e9renciation d\u00e9veloppementale des cellules souches dans et le long de la voie cellulaire OL (Fig. 2A). Cela a \u00e9t\u00e9 fait \u00e0 la PDMPP par Zhipeng You et Valerio Piscopo en utilisant la technologie CRISPR \/ Cas-9. Ces cellules vont maintenant \u00eatre tri\u00e9es (Fig. 2B) et ajout\u00e9es \u00e0 des plaques sp\u00e9ciales contenant des nanofibres afin de suivre leur potentiel de my\u00e9linisation (Fig. 2C).<\/p>\n

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Figure 2.<\/strong> Production de la lign\u00e9e iPSC SOX10-mOrange reporter. A) Image au microscope des OPC SOX10 positifs, montrant la fluorescence rouge; B) Les cellules sont tri\u00e9es en direct pour l\u2019expression du rapporteur et C) ensemenc\u00e9es sur des plaques de nanofibres pour \u00e9valuer la my\u00e9linisation, repr\u00e9sent\u00e9e en vert par l\u2019expression de la prot\u00e9ine de base de la my\u00e9line (MBP).<\/p><\/div>\n

 <\/p>\n

R\u00e9f\u00e9rences<\/em><\/h3>\n

1. He\u00df K, Starost LJ, Kieran NW, Thomas C, Vincenten MCJ, Antel J, Martino G, Huitinga I, Healy L, Kuhlmann T. Lesion stage dependent causes for impaired remyelination in MS. Acta Neuropathol- 2020 Jul 24.<\/em>
\n2. Franklin RJ, Ffrench-Constant C. Remyelination in the CNS: from biology to therapy. Nat Rev Neurosci. 2008;9(11):839-855. doi:10.1038\/nrn2480<\/em>
\n3. Luo JXX, Cui QL, Yaqubi M, Hall JA, Dudley R, Srour M, Addour N, Jamann H, Larochelle C, Blain M, Healy LM, Stratton JA, Sonnen JA, Kennedy TE, Antel JP. Human oligodendrocyte myelination potential; relation to age and differentiation. Ann Neurol. 2022 Feb;91(2):178-191. doi: 10.1002\/ana.26288.Epub 2022 Jan 10.<\/em>
\n4. Takahashi K, Tanabe K, Ohnuki M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131(5):861-872. doi:10.1016\/j.cell.2007.11.019<\/em>
\n5. Douvaras P, Fossati V. Generation and isolation of oligodendrocyte progenitor cells from human pluripotent stem cells. Nat Protoc. 2015;10(8):1143-1154. doi:10.1038\/nprot.2015.075<\/em>
\n6. Larochelle C , Wasser B, Jamann H, L\u00f6ffel JT, Cui QL, Tastet O, Stratton J, Schillner M, Luchtman D, Birkenstock J, Stroh A, Antel J, Bittner S, and Zipp F. Pro-inflammatory T-helper 17 directly harm oligodendrocytes in neuroinflammation. Proc Natl Acad Sci- 2021 Aug 24;118(34): e2025813118<\/em>
\n7. Jamann H, Cui Q, Desu HL, Pernin F, Tastet O, Halaweh A, Farzam-kia N, Mamane VH, Ou\u00e9draogo O, Cleret-Buhot A, Daigneault A, Lemaitre F, Arbour N, Antel J, Stratton J, Larochelle C \u2013 Contact-dependent granzyme B-mediated cytotoxicity of Th17 cells towards human oligodendrocytes – Frontiers Immunology \u2013 in press<\/em>
\n8. 8.Moore CS, fi Q, Warsi NM, Durafourt BA, Zorko N, Owen DR, Antel JP, Bar-Or A. Direct and Indirect Effects of Immune and Central Nervous System-Resident Cells on Human Oligodendrocyte Progenitor Cell Differentiation. J Immunol. 2015 Jan 15;194(2):761-72.<\/em>
\n9. Pernin F, JXX, Cui QL, Blain M, Fernandes MGF, Yaqubi M, Srour M, Hall J, Dudley R, Jamann H, Larochelle C, Zandee SEJ, Prat A, Stratton J, Kennedy TEK, Antel JP. Diverse injury responses of human oligodendrocyte to mediators implicated in multiple sclerosis -BRAIN \u2013 in press<\/em>
\n10. Starost L, Lindner M, Herold M, Xu YKT, Drexler HCA, He\u00df K, Ehrlich M, Ottoboni L, Ruffini F, Stehling M, R\u00f6pke A, Thomas C, Sch\u00f6ler HR, Antel J, Winkler J, Martino G, Klotz L, Kuhlmann T Extrinsic immune cell-derived, but not intrinsic oligodendroglial factors contribute to oligodendroglial differentiation block in multiple sclerosis. Acta Neuropathol. 2020 Sep 7. <\/em>
\n11. Lisak RP, Benjamins JA, Nedelkoska L, Barger JL, Ragheb S, Fan B, Ouamara N, Johnson TA, Rajasekharan S, Bar-Or A. Secretory products of multiple sclerosis B cells are cytotoxic to oligodendroglia in vitro. J Neuroimmunol. 2012 May 15;246(1-2):85-95.<\/em><\/p>\n

 <\/p>\n","protected":false},"excerpt":{"rendered":"

Chaque mois, dans les MRM Insights, un membre du R\u00e9seau MRM \u00e9crit sur les cellules…<\/p>\n","protected":false},"author":6,"featured_media":7937,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_bbp_topic_count":0,"_bbp_reply_count":0,"_bbp_total_topic_count":0,"_bbp_total_reply_count":0,"_bbp_voice_count":0,"_bbp_anonymous_reply_count":0,"_bbp_topic_count_hidden":0,"_bbp_reply_count_hidden":0,"_bbp_forum_subforum_count":0,"ngg_post_thumbnail":0,"footnotes":""},"categories":[162],"tags":[164,178,568,569,570],"class_list":["post-7976","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-mrm-insights-fr","tag-mrm-fr","tag-mrm-insights-fr","tag-multiple-sclerosis-fr","tag-oligodendrocytes-fr","tag-sclerose-en-plaques"],"acf":[],"yoast_head":"\nMRM Insights : Oligodendrocytes d\u00e9riv\u00e9s d\u2019IPSC \u2013 un mod\u00e8le de potentiel de 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