matrixNLRP3ASCASC speckCASP1 FLCasp 1 clactivityIL-1βIL-18GSDMD FL (53kDa)Cl 30kDapaSyn
Plasma Detectable, but BLQ ~410 pg/ml : 2 fold dilution LOD~1.6 pg/ml; limited spike recovery good NOMID gof >100X increase Barely measurable, dilutional linearity unknownk but team decision was to de-prioritize so no further work is planned could be run at risk BLOD (<7 pg/ml), Normal endogenous level unknownYes 1-2 ng/ml (1/10:10ul) 10ul 있어 본다는 거네 NOMID gof ~76X increase Planned: limited spike recovery, Assay available (No pooling required)Some total activity in plasma, but little/no inhibition with caspase-1 inhibitor (MSD S plex: LOD ~0.1 pg/ml) signal from recombinant caspase-1 is completely supressed when spiked into plasma, (but not CSF detectable but not strictly quantitative, as they are mostly between the LOD and the LLOQ So we need to keep in mind that comparison baseline may be more relative measurement)Endog level: ~0.2-0.8 pg/ml (TSD) 아니네: NOMID gof ~30X increase Planned: limited spike recovery Assay available (No pooling required) Required volume: 25ulEndog level: Pg/ml lequ ↑ >76x ≥130x
Plasma — NOMID mouse>↑100x>↑76x
20220922 Paul: share with Katy the ASC antibody details
Human / CSF
Assay: HumanX (→ s-plex, quanterix)O (Homebrewed MSD)XXXfeasible in 10X conc.X (→ S-plex by LSI)O (Olink target48)X (→ proteina: NTD)Cl 30kDa (아마 NM 22kDa)
Assay: mouse Bz-ATP MouseNot yetO (1/4, ~4 mice, pph)Testing feasibilityMSD S-PLEX (BLOD)Just measurable @4-fold diln; ~40 pg/ml (requires 30ul; ~6 mice) (hemo)Not detectible=NMNM
Atuka Tx effect? SNNA=
Atuka 2nd Tx effect?=↓ (098 & 660)↓ (098 & 660)==Slightly ↑
BzATP Mouse / model itself↑ (1.8x)↑ (monomer, 22kDa)= (mature)
098 3 mpk= (only trend ↓)↓ (100% normalization, 22kDa)↓ (mature, 22kDa) (note: FL is 31-35kDa)
098 30 mpk↓ trend (d=0.2)
660 10 mpk↓ trend (d=0.2)↓ (missed p)=↓ (missed p)↓ (missed p)=↑ trend
660 30 mpk↓ (2.4x)↓ trend (-20%, d=0.31))↓ (1.3x)↓ (monomer 22kDa)= (mature)↑ trend
BzATP Rat / Model itself↑ (1.3x)↑ (1.3x)↑ (2.8x)↑ (1.6x)↑ (1.4x)↑ (1.8x)
660 10 mpk?↑ (over measured?)↓ (2.5x)↓ (1.4-6x)== (mature form,↑ (1.3x, SN; ↓ (FL); cleaved GSDMD only one mouse)
660 30 mpkongoing
OHDA PS BzATP rat / 5d model itself↑ (2.5x)↑ (x7 SN)= (just missed) (d=0.6)↑ (1.8x)
9d model itself↑ (2.4x)
5d model itself (TH)
9d model itself (TH)trend ↑
Sortwell AAV-aSyn rat itself↑ (1.8x, Str)cleaved 4 & full-length↑ (cleaved & full-length, SN)↑ (1.8X, Str)
6-OHDA mouse
Human brain↑ (monomer 15-25 kDa & dimer 30-45kDa)↑ (FL)cleaved (17kDa)↑ / ↓ (FL)Cleaved GSDMD detected??
Human CSF Proteina↓ trendBelow LLOQ → accelerate?)Katy's probe-based assayBarely detectedHorrible (cl 일것)
CBID transfer Needed

20240312: Needed volume: NFL (3 µL), Casp1 activity assay (16 µL), total Casp1 ELISA assay (10 µL), and IL-18 assay (25 µL). If all the assays are conducted, ~54 µL of CSF needed. CSF volume is limited, we will discuss the priority of assays. Currently, priority order would be IL-18 > Casp1 activity assay > NFL > total Casp1 (detect Casp1 activity, we may need to check the Casp1 expression itself though difference was observed)

Atuka aSyn / BzATP rat dose grid

131030
Atuka aSyn mouse098098
BzATP rat660660
Atuka aSyn rat980MED

098: 2ND ATUKA mouse, 10 mpk, (vs AAV+veh) TH ↓ Putamen / NBB / Imperial

10 mpk (vs AAV+veh)30 mpk3 mpk1 mpk Vs veh% correctiondPost power, n=18, 1-sided
Alpha=0.05
098: 2ND ATUKA mouse↑ TH cell count SN48%0.9688%
↑ TH (wb) Striatum68%0.6965%
↓ TSPO striatum0.1>100%
Striatum Cleaved Caspase-1 (p20)100%1.0291%
Striatum IL-1b37%78%
660BzATP rat30 mpkVs vehStriatum Cleaved Caspase-1 (p20)61%29%
10 mpkVs vehStriatum Cleaved Caspase-1 (p20)96% (MED)51%
9801 mpkVs vehStriatum Cleaved Caspase-1 (p20)92% (MED)39%
1PutamenCSFCSFCross with other brain BM?
NBB14↑ (BUT Too many zero in 1 ratm)Correlation (only with) putamen, r=0.65 (all)SN GSDMD (0.43, pd), Put ASC dimer (0.51, all),
Imperial= (some high values in control)No correlation, but too many zero in putamen
Human= (onlink & somascan)

Uncertain Spans

locationtranscriptionuncertainty
Casp 1 cl Plasma cellEndog level: ~0.2-0.8 pg/ml (TSD) 아니네:The trailing Korean 아니네: and inline parenthetical context are partly cut at the cell boundary; preserved as visible.
Sortwell rowcleaved 4 & full-lengthcleaved 4 may be cleaved & due to OCR-confused & glyph; preserved as visible.
BzATP rat 660 30 mpkrow entirely marked ongoingCell shows only the word ongoing spanning multiple readout columns; rendered as colspan.
right-side power table sample sizesNBB 1 / 4Putamen sample sizes appear handwritten 1 / 4; reading is uncertain because the small font is partly faded.