Cografting astrocytes improves cell therapeutic outcomes in a Parkinson’s disease model

Abstract

In this study, we have attempted to exploit the neurotrophic actions of astrocytes and Nurr1+Foxa2 functions in this cell type to improve the therapeutic outcomes of NPC transplantation using an animal model of PD. Our in vitro assays using a coculture system and conditioned media treatment revealed that astrocytes, especially those cultured from the ventral midbrain (VM), where HNF3β) is a potent cofactor that synergizes the Nurr1-mediated antiinflammatory roles in glia (16). Furthermore, forced coexpression of Nurr1 and Foxa2 (Nurr1+Foxa2) in glia promotes the synthesis and release of various neurotrophic factors, indicating that engineering of Nurr1 and Foxa2 in glia could become a powerful strategy for treating CNS disorders. Representative images of TH + DA neurons differentiated from VM-NPCs in the presence of Ctx-NPCs (control), Ctx-Ast, or VM-Ast. Scale bar: 100 μm. Insets, enlarged images of the boxed areas (original magnification, ×400). (C) DA neuronal yields at differentiation day 6 (D6). (D-H) Morphometric measurement of DA neuron maturity assessed by neurite outgrowth lengths (D), soma size (E), and by the Sholl test (F and G). In the Sholl analysis, the total number of neurite crossings was counted in each circle with the radius increasing in steps of 15 μm (F). The critical value (G) is the radius at which there was a maximum number of neurite crossings. *P < 0.05, significantly different from the control; # P < 0.05, significantly different from Ctx-Ast, 1-way ANOVA. n = 3-5 wells (C) and 100 (D and E) and 25-30 (F and G) TH + cells in each group.(H) Representative TH + neuronal morphologies reconstructed in 3D by Neurolucida. (I-L) Synaptic densities on TH + fibers. (I) Representative confocal images of synapsin + TH + fibers. Scale bar: 25 μm. Puncta positive for the synaptic vesicle-specific markers SV2 (J), synapsin (K), and bassoon (L) on TH + fibers were counted. *P < 0.05, significantly different from the control; # P < 0.05, significantly different from Ctx-Ast. n = 20 TH + fibers for each group. (M) Presynaptic DA release. n = 3 independent cultures. DA levels were measured in the media conditioned in the differentiated cultures for 24 hours (D13-D15) and in cultures evoked by KCl-mediated depolarization for 30 minutes. *P < 0.05; # P < 0.05, 1-way ANOVA. The Journal of Clinical Investigation R E S E A R C H A R T I C L E4 6 5 jci.org Volume 128 Number 1 January 2018 (Supplemental Figure 1B; supplemental material available online with this article; https://doi.org/10.1172/JCI93924DS1), immunocytochemical analyses revealed that major cell populations in the Ctx-and VM-astroglial cultures at day in vitro (DIV) 14 were positive for GFAP (85% and 82%), CD44 (79% and 76 %), GLT-1 (58% and 71 %), and GLAST protein (71% and 77%), respectively (Supplemental Figure 1A), except S100β (20% and 32%), a soluble protein associated with harmful reactivation of astrocytes (22). A few (0.24% ± 0.35% in Ctx-Ast and 0.43% ± 0.39% in VM-Ast, where Ast indicates astrocytes) and none of the cells in these c...

Boosting Antioxidant Self-defenses by Grafting Astrocytes Rejuvenates the Aged Microenvironment and Mitigates Nigrostriatal Toxicity in Parkinsonian Brain via an Nrf2-Driven Wnt/β-Catenin Prosurvival Axis

“…Based on this background, VM-As appear uniquely positioned to drive neuroprotective and regenerative programs in PD, and recent preclinical studies used co-transplantation of As to promote the regenerative effects of co-transplanted stem cells in rodent PD models (Yang et al, 2014;Song et al, 2018). However, how the aged brain responds to pharmacological or cellular therapeutical interventions is not well documented.…”

Transplantation of Nurr1‐overexpressing neural stem cells and microglia for treating parkinsonian rats

“…Moreover, this study found that most implanted cells were localized in the transplant area, and only a few of them were observed in the striatum of PD rats in the NNSC + NMG group after months (Figure ). Based on the results and previous studies, the molecular mechanisms underlying the neuroprotective effects of Nurr1 on PD included the reduction of inflammatory factors and the secretion of neurotrophic factors and other cytokines, such as SHH and FGF8 …”

“…Importantly, the key obstacles to the success of preclinical trials in PD are the poor survival of grafted cells . After cell transplantation, the hostile host brain environment is mainly responsible for unsatisfactory results . A few studies targeted the host brain to improve therapeutic effects .…”

Parkinson-Erkrankung und Neuroinflammation – Zelluläre Pathologie, Mechanismen und therapeutische Optionen

“…B. kontinuierlich neurotrophe Faktoren zu sezernieren, wie erfolgreich in Tiermodellen gezeigt werden konnte [20,105]. Der neurorestaurative Effekt nach Transplantation von neuronalen Vorläuferzellen in das Striatum wird durch begleitende Transplantation von Astrozyten deutlich verbessert [106]. Noch eleganter ist ein Ansatz, der eine Zelltransplantation vermeidet und Astrozyten direkt im Striatum in situ mittels stereotaktisch applizierter lentiviraler Gen-Vektoren zu dopaminergen Zellen umwandelt, damit sie die Funktion der degenerierten dopaminergen Neurone übernehmen [107].…”