Three major αsyn domains (N-terminal / NAC / C-terminal acidic tail), Synapse and aSyn (Calleri 2015 90% presynaptic deposits), TM for aSyn programs (Monomer / Pansson 2018 fibril CDMS, αSyn oligomer enrichment plan, paSyn / Fragments / LB / PD vs MSA / DLB SPAL? rows)

Three major αsyn domains

Three major domains	Aa N-terminal domain	central domain α-syn amyloid component (NAC)	C-terminal acidic tail
	1-60 (or 1-67)	aa 61-95	aa 96-145
	Consists of five imperfect repeats, each 11 amino acids in length, with the consensus sequence KTLEKEUSeoSeo
Function	binds lipids and adopts alpha helical structures DOI: 10.13140/RG.2.2.35950.97602	Hydrophobic, involved in aggregation	accountable for most interactions with other proteins and small molecules

over half of which consists of seven imperfect repeats with the consensus sequence KTKLGV (residues 7-87)

Synapse and aSyn

(Calleri, 2015 #1185 review): In PD brain, 90% or even more of α-syn aggregates in the brain of PD patients are located in the presynapses in the form of very small deposits. In parallel, dendritic spines are retracted, whereas the presynapses are relatively preserved, suggesting that neurotransmitter deprivation is an early event in PD pathogenesis.

TM (target modulation) for aSyn programs

What to target	consider	TAK-341	BM	SNCA RTV in 변?
Monomer	Native aSyn is monomeric and helical oligomers; Available
	CSF total aSyn / Plasma	Affinity for monomeric recombinant αSYN: in cell pSyn (Otex stained) and 75 pM (solution phase)	MSD 일감 BoBe iv 1-1 1-95 MSD ELISA	MSD ELISA
oligomers	(Pansson, 2018 #2075) study with recombinant fibril (in buffer) Type 1 Trimedi fiber Type 2 Triangular ribbon stainflakes Mass um height length 5.7 - 5 - 930 μm 167.8 MDa lower charge CD: ~85 kDa, 9900 αSyn molecules, average length 0.9 µM	Affinity for aggregated αSYN: in cell ~25 ~930 pM (not substantially different than monomers)	we (and Roche) have not been able to measure CSF oligomeric αSyn species; (Olga) CDMS [charge detection mass spectrometry] single particle technique can be used to characterize multimeric species (αsyn or tau oligomers and fibrils) - to distinguish or quantify different species (polymorphic structures). Charge detection mass spec to profile different species of α-syn enclosed on size and charge. (Olga 가 LCMS 1, HV/PD/MSA CSF 비교 profiling 한 (1+1 11 slides 'Hu 미국 일컬), 2 brain 1 / @ 02-1F2)
20231116: successfully detected size distribution of recombinant fibrils of α-syn, work to assess feasibility of detecting multimers in biofluid is still ongoing. Goal: current efforts in biofluid rely on enrichment with specific antibodies (eg TAK-341). Optimal reagents for enrichment will need to be identified if feasibility looks promising. Pilot experiment Reproduced Pansson (more difference?) starts due to different methods? Initial feasibility testing result by end Jan, success means detecting (and quantifying to some extent) endogenous αSyn oligomers (~8 of fal) another experiment To test biased antibodies to recruit; Working with CSF Matrix others 2/13 (full); CSF study (Otake-san) 안 일컬 02 (이 CSF) 02-1F2 (Otake) / preclinical 외 그 도와 가는 'all' clinical (1) (1)) 'rsay' / development '5' No more working with Waters (피 Olga 9.8) NOTE: Brain validation 외 외 '5'-5/12 외 '미음'; 'disease' 'stage'? '5' 외 '실 외 '비외 '?'; 1. Disease severity correlation 이 가 이 . 20240210 The effort is covered by the indirect BST budget as part of evaluating CDMS technology for multiple analytical applications. Successful pilot experiments with recombinant α-syn (PFFs) and α-synuclein completed in 2023, demonstrating detection of large molecular weight species in the mixture of fibrils Outstanding questions Does CDMS have the sensitivity to detect very low amounts of endogenous α-syn oligomers in biofluids - currently being tested using CSF Two different populations of (αs recombinant species, detected (small oligomers and larger fibrils)) TAK A-SYN Current feasibility testing Initial feasibility run is ongoing (readout expected Feb2024) using several anti-α-synuclein antibodies for enrichment of α-syn oligomers and PD CSF Depending on the outcome of the initial feasibility run, additional feasibility runs may be performed using alternative anti-α-synuclein antibodies and additional PD CSF samples (Q2 2024) We will also be developing LCMS to look at fragments of α-syn and pS129
paSyn / Fragments / LB	Bind in PM (else IB?)
As we do not currently know the identity of the toxic species of α-syn, it is difficult to predict the outcome of treatment with such compounds, which may inadvertently result in an increase in the toxic species; Fibrillar dissociation could, for example, lead to oligomers a-syn release and its downstream effects. deoriumental effects oligomers & fibrillar 다 일감 어 가 다.

PD vs MSA / DLB ? (SPAL?)

	content
MSA	Cell to cell generating in PD occurs via the neuronal connectome. αSYN in MSA is mainly localized to oligodendrocytes. The oligodendrocyte connectome is nearly identical to the neuronal connectome (discussed with Alain Dagher and others at the MJFF αSYN conference). αSYN filament structure is different between PD, MSA and DLB (Schweinghouse, 2020 #2960). This risk was mitigated by AZ showing that MEDI-1141 can bind to MSA aggregates.
	1 Seeding capacity, 1 Of A-syn vs DLB-aSyn (Prosinier 2015 #2965; Peng 2018 #2961)
	1 Exclusively αSYN pathology in about 62% cases MSA vs 25% in PD (Kovacs ‘15; Campos 2021 #2960) (대 다 가 마 가 다 일컬 시 나오네); Fast progressing

(Stephens et al., 2019 #1785) there was a greater Fluid; How much to local? TAK-341 in hippo → ↓ Syn-1 in ipsi hippo, ipsi hippocampal axon -왼쪽의 downstream