Identifying secreted biomarkers of dopaminergic ventral midbrain progenitor cells
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Identifying secreted biomarkers of dopaminergic ventral midbrain progenitor cells. / Rifes, Pedro; Isaksson, Marc; Rusimbi, Charlotte; Ramón Santonja, Adrián; Nelander, Jenny; Laurell, Thomas; Kirkeby, Agnete.
In: Stem Cell Research and Therapy, Vol. 14, No. 1, 354, 2023.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Identifying secreted biomarkers of dopaminergic ventral midbrain progenitor cells
AU - Rifes, Pedro
AU - Isaksson, Marc
AU - Rusimbi, Charlotte
AU - Ramón Santonja, Adrián
AU - Nelander, Jenny
AU - Laurell, Thomas
AU - Kirkeby, Agnete
N1 - Publisher Copyright: © 2023, The Author(s).
PY - 2023
Y1 - 2023
N2 - Background: Ventral midbrain (VM) dopaminergic progenitor cells derived from human pluripotent stem cells have the potential to replace endogenously lost dopamine neurons and are currently in preclinical and clinical development for treatment of Parkinson’s Disease (PD). However, one main challenge in the quality control of the cells is that rostral and caudal VM progenitors are extremely similar transcriptionally though only the caudal VM cells give rise to dopaminergic (DA) neurons with functionality relevant for cell replacement in PD. Therefore, it is critical to develop assays which can rapidly and reliably discriminate rostral from caudal VM cells during clinical manufacturing. Methods: We performed shotgun proteomics on cell culture supernatants from rostral and caudal VM progenitor cells to search for novel secreted biomarkers specific to DA progenitors from the caudal VM. Key hits were validated by qRT-PCR and ELISA. Results: We identified and validated novel secreted markers enriched in caudal VM progenitor cultures (CPE, LGI1 and PDGFC), and found these markers to correlate strongly with the expression of EN1, which is a predictive marker for successful graft outcome in DA cell transplantation products. Other markers (CNTN2 and CORIN) were found to conversely be enriched in the non-dopaminergic rostral VM cultures. Key novel ELISA markers were further validated on supernatant samples from GMP-manufactured caudal VM batches. Conclusion: As a non-invasive in-process quality control test for predicting correctly patterned batches of caudal VM DA cells during clinical manufacturing, we propose a dual ELISA panel measuring LGI1/CORIN ratios around day 16 of differentiation.
AB - Background: Ventral midbrain (VM) dopaminergic progenitor cells derived from human pluripotent stem cells have the potential to replace endogenously lost dopamine neurons and are currently in preclinical and clinical development for treatment of Parkinson’s Disease (PD). However, one main challenge in the quality control of the cells is that rostral and caudal VM progenitors are extremely similar transcriptionally though only the caudal VM cells give rise to dopaminergic (DA) neurons with functionality relevant for cell replacement in PD. Therefore, it is critical to develop assays which can rapidly and reliably discriminate rostral from caudal VM cells during clinical manufacturing. Methods: We performed shotgun proteomics on cell culture supernatants from rostral and caudal VM progenitor cells to search for novel secreted biomarkers specific to DA progenitors from the caudal VM. Key hits were validated by qRT-PCR and ELISA. Results: We identified and validated novel secreted markers enriched in caudal VM progenitor cultures (CPE, LGI1 and PDGFC), and found these markers to correlate strongly with the expression of EN1, which is a predictive marker for successful graft outcome in DA cell transplantation products. Other markers (CNTN2 and CORIN) were found to conversely be enriched in the non-dopaminergic rostral VM cultures. Key novel ELISA markers were further validated on supernatant samples from GMP-manufactured caudal VM batches. Conclusion: As a non-invasive in-process quality control test for predicting correctly patterned batches of caudal VM DA cells during clinical manufacturing, we propose a dual ELISA panel measuring LGI1/CORIN ratios around day 16 of differentiation.
KW - Biomarkers
KW - Cell replacement therapy
KW - Dopamine progenitors
KW - ELISA
KW - Extracellular vesicles
KW - Mass spectrometry
KW - Parkinson’s disease
KW - Quality control
KW - Secretome
U2 - 10.1186/s13287-023-03580-5
DO - 10.1186/s13287-023-03580-5
M3 - Journal article
C2 - 38072935
AN - SCOPUS:85179366270
VL - 14
JO - Stem Cell Research & Therapy
JF - Stem Cell Research & Therapy
SN - 1757-6512
IS - 1
M1 - 354
ER -
ID: 377803854