Abstract
Degeneration of dopamine (DA) neurons in the midbrain underlies the pathogenesis of Parkinson’s disease (PD). Supplement of DA via L-DOPA alleviates motor symptoms but does not prevent the progressive loss of DA neurons. A large body of experimental studies, including those in nonhuman primates, demonstrates that transplantation of fetal mesencephalic tissues improves motor symptoms in animals, which culminated in open-label and double-blinded clinical trials of fetal tissue transplantation for PD1. Unfortunately, the outcomes are mixed, primarily due to the undefined and unstandardized donor tissues1,2. Generation of induced pluripotent stem cells enables standardized and autologous transplantation therapy for PD. However, its efficacy, especially in primates, remains unclear. Here we show that over a 2-year period without immunosuppression, PD monkeys receiving autologous, but not allogenic, transplantation exhibited recovery from motor and depressive signs. These behavioral improvements were accompanied by robust grafts with extensive DA neuron axon growth as well as strong DA activity in positron emission tomography (PET). Mathematical modeling reveals correlations between the number of surviving DA neurons with PET signal intensity and behavior recovery regardless autologous or allogeneic transplant, suggesting a predictive power of PET and motor behaviors for surviving DA neuron number.
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All requests for raw and analyzed data and materials will be promptly reviewed by the corresponding author and the University of Wisconsin–Madison to verify whether the request is subject to any intellectual property or confidentiality obligations. Any data and materials that can be shared will be released via a material transfer agreement.
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Acknowledgements
This research was supported by grants from the National Institutes of Health–National Institute of Neurological Disorders and Stroke (NS076352, NS096282 and NS086604), the Eunice Kennedy Shriver National Institute of Child Health and Human Development (U54 HD090256), P51OD011106, the National Medical Research Council of Singapore (MOH-000212 and MOH-000207), the Dr. Ralph & Marian Falk Medical Research Trust, the University of Wisconsin–Madison Office of Vice Chancellor for Research and Graduate Education, the Cellular and Molecular Pathology Graduate Program, the Neuroscience Training Program and the Departments of Radiology and Medical Physics at the University of Wisconsin–Madison. This project was possible due to the dedication and support of Wisconsin National Primate Research Center veterinarians and animal care technicians, especially C. Boettcher, K. Fuchs and D. Schalk. We are grateful to P. Perez Toro, S. Brady, K. MacManus, A. Payne and L. Fox for facilitating behavioral testing procedures during their undergraduate studies.
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Contributions
Y.T. reprogrammed monkey iPSCs, performed the cell culture, DA differentiation, immunostaining, transplantation, data analysis and interpretation and wrote the manuscript. S.V., K.B. and J.M. performed MPTP post-surgical care and cell transplantation. M.Z. and J.H. produced PET images and related analysis. J.L. and L.Y. reprogrammed the monkey iPSC and performed the cell culture. M.O. created the real-time intraoperative MRI targeting roadmaps and PET–MRI co-registrations. Y.C. constructed the GFP lentivirus plasmid. S.P. and N.S. collected and analyzed behavioral data and performed histological evaluations. V.B. performed immunohistochemistry. W.B. analyzed real-time intraoperative MRI targeting roadmaps. T.B. produced [11C]DTBZ. H.A.S. performed necropsies and related data interpretation. B.C. performed analysis and interpretation of PET data. M.E. conceived and designed the experiments, performed intracarotid MPTP, performed cell transplantation and animal evaluations, data analysis and interpretation and wrote the manuscript. S.-C.Z. conceived and designed the experiments, data analysis and interpretation and wrote the manuscript.
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S.-C.Z. is a cofounder of BrainXell, Inc.
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Extended data
Extended Data Fig. 1 DA neuron generation and MPTP PD model.
a, Representative images of pluripotent stem cell marker expression in iPSCs generated from rhesus macaque fibroblasts. b,c, Representative images of mDA progenitor marker (b) and DA neuron marker (c) in differentiating cells from rhesus macaque iPSCs. Scale bar: 50μm. Data are representative of at least 5 independent experiments (a-c). d, Images of TH immunostaining in the substantia nigra from allogenic and autologous rhesus monkeys. e, Stereological quantification of TH+ neurons in the substantia nigra of allogenic and autologous rhesus monkeys. f, Percentage of TH+ cell reduction in the MPTP-treated substantia nigra compared to the unlesioned side. The data are presented as mean ± s.d. (n = 5 biologically independent monkeys in each group) in e, f.
Extended Data Fig. 2 Mood behavior in transplanted monkeys.
a, The anxious pacing (AP) behavior observed in monkeys receiving allogenic or autologous transplantation from 12 months before transplantation to 24 months after transplantation. The transplantation happened at month 0. Lines show mean values for every 6 months from the allogenic group or the autologous group. b, The lack of motivation (LOM) behavior observed in monkeys receiving allogenic or autologous transplantation from 12 months before transplantation to 24 months after transplantation. c, The self-injury behavior (SIB) observed in monkeys receiving allogenic or autologous transplantation from 12 months before transplantation to 24 months after transplantation.
Extended Data Fig. 3 Graft evaluation in vivo.
a,b, Quantification of [11C]DTBZ graft binding potential in contralateral (untreated) putamen (a) and caudate (b) from allogenic and autologous monkeys before and after transplantation. The data is presented as mean ± s.d. (n = 4 per group). c,d, Quantification of the volume of uptake in contralateral (untreated) putamen (c) and contralateral caudate (d) from allogenic and autologous monkeys before and after transplantation. The data is presented as mean ± s.d. (n = 4 per group).
Extended Data Fig. 4 Overview of the graft.
a, Representative images of GFP immunostaining and Nissl staining in brain sections of allogenic and autologous animals. The red arrows point to the grafts. b, H&E staining in brain sections of allogenic and autologous animal. Enlarged images correspond to the yellow area in the respective grafts. All grafts (if present) in monkeys from both groups were examined. Data are representative of at least 3 sections having grafts from each monkey.
Extended Data Fig. 5 Histological analysis of graft.
a, Representative images of TH immunostaining in brain sections of allogenic and autologous animals. Enlarged images correspond to the grafts. b, Representative images of TH+ fiber extension area in control and MPTP brain hemisphere. c, TH immunostaining in the putamen from MPTP lesion side and unlesioned side. Scale bar: 10 µm. All grafts (if present) in monkeys from both groups were examined. Data are representative of at least 3 sections having grafts from each monkey (a-c).
Extended Data Fig. 6 Caudate graft in autologous monkeys.
Representative image of TH immunostaining in autologous monkey caudate region. The inset area is enlarged below. All grafts (if present) in monkeys from both groups were examined. Data are representative of at least 3 sections having grafts from each monkey.
Extended Data Fig. 7 Cellular composition in grafts.
a, Representative images of TH and GIRK2 or Calbindin immunostaining in grafts. Scale bars: 50 μm. b, Representative images of vGLUT1, 5-HT and GABA immunostaining in grafts. Scale bars: 50 μm. c, Representative images of COL1A1 immunostaining in and outside of grafts. scale bars: 50 μm. The white dash lines mark the edge of the graft. All grafts (if present) in monkeys from both groups were examined. Data are representative of at least 3 sections having grafts from each monkey (a-c).
Extended Data Fig. 8 Immune response evaluation in grafts.
a, Histological analysis of T cells (CD3 and CD45), microglia (CD68) and astrocyte (GFAP) marker in grafts from allogenic and autologous animals. Scale bar: 100 μm. b, Representative images of GFP and GFAP immunostaining in allogenic and autologous monkeys. Scale bar: 50 μm. The white dash lines mark the edge of the graft. All grafts (if present) in monkeys from both groups were examined. Data are representative of at least 3 sections having grafts from each monkey (a-b).
Extended Data Fig. 9 Regression analysis on the relation between DA neuron numbers and behavioral recovery/PET.
a, Linear regression analysis between ipsilateral caudate [11C]DTBZ binding potential and FMS. b, Linear regression analysis between ipsilateral caudate [11C]DTBZ binding potential and CRS. c, Linear regression analysis between ipsilateral caudate [11C]DTBZ binding potential and CRS recovery rate. d, Linear regression analysis between ipsilateral caudate [11C]DTBZ binding potential and surviving TH+ neuron numbers. e, Linear regression analysis between ipsilateral caudate [11C]DTBZ binding potential and caudate surviving TH+ neuron numbers. f, Linear regression and logistic fitting analysis of FMS and total surviving TH+ neuron numbers in grafts. The Pearson’s r, significance (p value) and R2 (coefficient of determination) were assessed by two-tailed Pearson’s correlation analysis in a-f.
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Tao, Y., Vermilyea, S.C., Zammit, M. et al. Autologous transplant therapy alleviates motor and depressive behaviors in parkinsonian monkeys. Nat Med 27, 632–639 (2021). https://doi.org/10.1038/s41591-021-01257-1
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DOI: https://doi.org/10.1038/s41591-021-01257-1
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