In vitro GBA-PD tail, 20191125 TAK Okai matrix, 20201209_ACi overview, Animal models of GBA-PD start
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In vitro GBA-PD tail, 20191125 TAK Okai matrix, 20201209_ACi overview, Animal models of GBA-PD start
In vitro GBA-PD (continued)
no.
assay / readout
model
observed / note
current / future plan
3
α-Syn release into culture sup. (MSD ELISA)
GBA-PD (N370S/wt) iPSC-DaNs from Oxford university
No increase was observed in α-Syn release into culture sup. Between WT and GBAL444P/L444P iPS-DaN Regarding to the cells from Oxford university, we have received only three iPSC lines due to their QCs (two healthy and one GBA-PD derived (N370S/wt).
Requesting other iPS lines to Oxford univ. Planning to start iPSC expansion and stock
4
ER stress (qRT-PCR)
GBA mut iPSC-DaNs
Increase of ER stress gene expression in some iPS-DaN clones was observed
Planning a replication experiments
5
DA oxidization
GBA mut iPSC-DaNs
On-going (collaboration with Kunisada G)
Assay development has been planned in Ando G in AXL (May and June)
Below from 20191125 TAK Okai san slide gba-gt slide
GBA perturbation
Sssss
Control
WES (Cell lysate)
MSD ELISA (Cell sup)
p-α-SYN WES (Cell lysate)
Native Western (Cell lysate)
SEC1 (Cell lysate)
WES (Cell pellet)
D409H
B7
No change
Not detected
No change
L444P
아래 ACi 에도 O
B7
N370S
B7
N370S (from Oxford univ.)
HV derived iPS
D409H +AAV-SNCA
B7 +AAV-SNCA
No change
Not detected
No change
SYN TRI cell + CBE
아래 ACi 에도 O
SYN TRI cell
L444P + PFF
B7 + PFF
N370S + PFF
B7 + PFF
N370S + PFF
Below from 20201209_ACi overview
In vitro models
αSyn pathology
Autophagy-lysosome pathway (ALP) abnormality
Accumulation
Phosphorylation
excretion
Gene expression
Protein expression
Cathepsin D activity
GBA L444P/L444P iPSC-DAn
위 TAK slide 에 O
No change
No change
SNCA triplication iPSC-DAn CRF
위 TAK slide 에 O
A53T SNCA DAn CRF
위 TAK slide 에 X
Animal models of GBA-PD (start)
Animal models of GBA-PD
(20200414 Tak) There was no difference in Sarkosyl-soluble or insoluble fraction of α-syn or p-α-syn accumulation (TH, WB),
(20200414 Tak) There was no difference in Sarkosyl-soluble or insoluble fraction of α-syn or p-α-syn accumulation (TH, WB)
20210405 ARNU Sugi) L444P had no effect on (insoluble) paSYN (contra Cortex, WB) in PFF mice