O4‐10‐01: Characterizing Cell Type‐specific Expression Signatures in Brain Using Single Cell Rna Sequencing

Patel, Tulsi; Carnwath, Troy; Lewis‐Tuffin, Laura J.; Allen, Mariet; Lincoln, Sarah; Wang, Xue; Deniz, Kaancan; İş, Özkan; Martin, Elizabeth; Garcia, César A.; Louie, Christopher; ReFaey, Karim; Durán, Iván; Guerrero-Cázares, Hugo; Wharen, Robert E.; Chaichana, Kaisorn L.; Quiñones‐Hinojosa, Alfredo; Ertekin-Taner, Nilüfer

doi:10.1016/j.jalz.2019.06.4794

Cited by 2 publications

(1 citation statement)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CSF1R signaling is known to be important for microglial proliferation in response to stress or injury, with increased M-CSF expression observed upon in vivo LPS treatment (56), upon ischemic injury (57), in Alzheimer's disease and its animal models (56,58,59), and in aged mice (56) and human (60). It can be hypothesized that CSF1R-mediated proliferation or functional changes of microglia are of increased importance in maintaining brain homeostasis during aging, explaining the late-onset of ALSP.…”

Section: Discussionmentioning

confidence: 99%

An adapted stem cell-derived microglia protocol for the study of microgliopathies and other neurological disorders

Dorion,

Casas,

Yaqubi

et al. 2023

Preprint

View full text Add to dashboard Cite

BackgroundAdult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is a primary microgliopathy caused by pathogenic variants in the colony-stimulating factor 1 receptor (CSF1R) gene. Since CSF1R signaling is crucial for microglia development, survival and function, induced pluripotent stem cell-derived microglia (iMGL) represent an excellent tool in studying microglial defects caused by ALSP patient-specificCSF1Rvariants.MethodsSerial modifications to an existing iMGL protocol were made, including but not limited to changes in growth factor combination to drive microglial differentiation, until successful derivation of microglia-like cells from an ALSP patient carrying a c.2350G > A (p.V784M)CSF1Rvariant. Using healthy control lines, the quality of the new iMGL protocol was validated through cell yield assessment, measurement of microglia marker expression, transcriptomic comparison to primary microglia, and evaluation of inflammatory and phagocytic activities. Similarly, molecular and functional characterization of the ALSP patient-derived iMGL was carried out in comparison to healthy control iMGL.ResultsThe newly devised protocol allowed the generation of iMGL with enhanced transcriptomic similarity to primary human microglia and with higher phagocytic and inflammatory competence at ∼3-fold greater yield compared to the original protocol. Using this protocol, decreased CSF1R autophosphorylation and cell surface expression was observed in iMGL derived from the ALSP patient compared to those derived from healthy controls. Additionally, ALSP patient-derived iMGL presented a migratory defect accompanying a temporal reduction in purinergic receptor P2Y12 (P2RY12) expression. Finally, ALSP patient-derived cells showed surprisingly high phagocytic capacity, which was associated with higher lysosomal content.ConclusionsWe optimized a pre-existing iMGL protocol, generating a powerful tool to study microglial involvement in human neurological diseases. Using the optimized protocol, we have generated for the first time iMGL from an ALSP patient carrying a pathogenicCSF1Rvariant, with preliminary characterization pointing toward functional alterations in migratory and phagocytic activities.

show abstract