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Identifying secreted biomarkers of dopaminergic ventral midbrain progenitor cells

Rifes, Pedro LU ; Isaksson, Marc LU ; Rusimbi, Charlotte ; Santoja, Adrian Ramón ; Wahlestedt, Jenny Nelander LU orcid ; Laurell, Thomas LU and Kirkeby, Agnete LU (2023) In Stem Cell Research & Therapy 14(1).
Abstract
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... (More)
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. (Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Stem Cell Research & Therapy
volume
14
issue
1
article number
354
pages
16 pages
publisher
BioMed Central (BMC)
external identifiers
  • pmid:38072935
  • scopus:85179366270
ISSN
1757-6512
DOI
10.1186/s13287-023-03580-5
language
English
LU publication?
yes
id
05f01a9e-a9e5-4aee-8a27-0e7d5967c929
date added to LUP
2023-11-24 13:10:01
date last changed
2024-01-04 10:57:57
@article{05f01a9e-a9e5-4aee-8a27-0e7d5967c929,
  abstract     = {{Background<br/>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.<br/><br/>Methods<br/>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.<br/><br/>Results<br/>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.<br/><br/>Conclusion<br/>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.}},
  author       = {{Rifes, Pedro and Isaksson, Marc and Rusimbi, Charlotte and Santoja, Adrian Ramón and Wahlestedt, Jenny Nelander and Laurell, Thomas and Kirkeby, Agnete}},
  issn         = {{1757-6512}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{BioMed Central (BMC)}},
  series       = {{Stem Cell Research & Therapy}},
  title        = {{Identifying secreted biomarkers of dopaminergic ventral midbrain progenitor cells}},
  url          = {{http://dx.doi.org/10.1186/s13287-023-03580-5}},
  doi          = {{10.1186/s13287-023-03580-5}},
  volume       = {{14}},
  year         = {{2023}},
}