Atuka NHP Results matrix tail, Woodruff lab a-syn of mouse, PFF model (mouse pre-formed α-synuclein fibril), TAKEDA w Atuka Lentivirus, Gordon 2018 #587 / Chu 2019 / Luk 2012 / Duffy 2018 / Patterson 2019
Atuka NHP Results matrix (continued)
| sub | sample | method | results | |
|---|---|---|---|---|
| NDGN | TH cell | SN | stereology | reduction of TH+ neurons; no spreading pathology |
| atrophy | Whole brain | 3T MRI | ||
| WM | DTI | |||
| a-syn | pSer129 aSyn | forebrain, midbrain, brainstem | qualitative histology | |
| Total aSYN (of primary interest for this study), aSYN oligomers, aSYN seed assays | CSF | |||
| others | t-Tau, p-Tau, Inflammatory BM Exosomal analytes |
Woodruff lab
| injection material | recipient | note | |
|---|---|---|---|
| Woodruff lab | a-syn of mouse | mouse | accelerated model: Motor deficits start around 1 months post PFF injection, (4 month required for further progression) |
| [기존모델] a-syn of human (synthetic), into mouse striatum, Motor deficits start around 4 months post PFF injection, (6-8 month required for further progression) | |||
아래는 여전히 기존 model같은데? [accelerated model 아니고] (Gordon, 2018 #585), Single injection of αSyn PFF into right dorsal striatum (8 ug in 2uL injected 0.2 uL/min) with NLRP3i MCC950 treatment via drinking water for 6 mo, starting from the day before PFF injection
| |||
PFF model (mouse, pre-formed α-synuclein fibril)
From Sci Transl Med. 2018; 10(465) (NLRP3 & PD paper from Woodruff lab) & Science 2012; 338: 949-953 (original report on mouse PFF model)
Method:
- Single injection of pre-formed α-synuclein fibril injection into right dorsal striatum (8 ug in 2uL, injected 0.2 uL/min)
- NLRP3i MCC950 treatment via drinking water for 6 mo, starting from the day before PFF injection
Observed pathology:
- NLRP3 activation (Day 30)
- Transmission of Lewy body pathology (deposition of hyper-phosphorylated serine 129 (pS129) α-synuclein in interconnected sites (cell-to-cell transmission) (Day 30-180)
- Progressive loss of dopaminergic neurons in the substantia nigra (-Day 180)
- Progressive deterioration on behavioral tests indicative of impaired motor coordination and balance without gross behavioral abnormality (-Day 180)
Figure panels:
- Distribution of pSyn accumulation after single PFF injection — PFF 30d / PFF 90d / PFF 180d coronal series
- TH neurons containing pSyn inclusions — panel K
- TH neuron quantification — TH
- Behavioral studies — D Rotarod / Wire hang / Open field activity; E Y maze / TST
- NLRP3 activation — L caspase-1 (p20) and ASC fold change PBS PFF (Day 30)
Series legend (behavioral): non-injected (wt) / PBS 30 d (wt) / PFF 30 d (wt) / PFF 90 d (wt) / PFF 180 d (wt) / PFF 180 d (Snca-/-)
Spreading models — TAKEDA w Atuka and key references
| injection material | recipient | results / notes | |
|---|---|---|---|
| TAKEDA w Atuka | Lentivirus + α-syn tg mice | In both non-WJ PLFH DQG α-syn tg mice |
• Goal: if, in the NHP model, TAK-341 is shown to reduce αSYN spreading and this reduction in spreading is predicted by suppression of CSF αSYN. • Method: TAK-341 (IDP) The contralateral non-injected hippocampus of non-tg mice and α-syn tg mice also displayed high levels of α-syn immunoreactivity, indicating that the LV-H[SUHVVHG α-syn had spread from the injected right hippocampus to the left hippocampus, contralateral non-injected hippocampus of non-tg mice and α-syn tg mice also displayed high levels of α-syn immunoreactivity, indicating that the H[SUHVVHG α-syn had spread from the injected right hippocampus to the left hippocampus • Timeline: (tak341 e POC doc updated 060619) the model will be validated by Q1 2020, and evaluation of TAK-341 in the model will be completed by Q1 2021, Human PFFs injected on March 26, 2020 when Suzhou facility reopened after COVID • Plasma, CSF, DAT/VMAT2 scans, and histopathology collected at 2 and 4 month post injection of PFFs |
| (Gordon, 2018 #587) | Mice 일것 | Good example! Read! | |
| (Chu, 2019 #591) | unilateral putamen injections of aSyn PFF synthesized from recombinant -syn | NHP cynomolgus Monkeys, 6 to 10 y of age |
• aSyn: 12-15 m after injection, phosphorylated a-syn in ipsilateral SN (not contralateral) and not adjacent to the injection site. a-Syn inclusions were thioflavin-S-positive, SN에도 감, cortex간 증거 없음. • neurodegeneration: ipsilateral SN with significant reduction (29.30%) of DA neurons • DATscan: 오히려 ↑ (upregulation 이라네) | baseline | 3m | 6m | 9m | 12m | 15m | |
| (Luk, 2012 #612) | WT aSyn PFF, & PFFs assembled from human a-Syn1-120Myc, a C-terminal truncated form of WT human aSyn containing Myc epitope | Mice, injection into the dorsal neostriatum | 30일에, Cortex 등 far away region 까지 p-aSyn 감. (90일에 더 심함), 둘다 멀리 갔는데, WT aSyn 보 주입시에 더 많이 감. |
| (Duffy, 2018 #1369) (Sortwell's lab) | aSyn PFF (recombinant but generated from mouse aSyn), Striatum (unilateral), two injection sites at the same tie. | (Male Fischer 344) rats (n=114) |
- 1m post injection: in SN: among paSYN-positive inclusions, i) 88.2% had either oligomeric conformation. ii) 56.4% had fibrillar aggregates, iii) 31.7% had oligomeric conformation. iv) many (?) had thioflavins-s-positive. - 2m post injection: in SN: paSYN is peak (fig4d) (in bilateral cortex, ipsilateral SNc), MHC-I reactive microglial morphology (ie ↑ soma area) 즉, paSyn 과 microglial pathology 는 i) 같이 가고 ii) loss 에 선행한다! - 5-6m post injection, widespread SN (BY 35%) - Patterson 2nd) 이 실험에서 neurodegeneration 만 ipsilateral 보였는데 SN d contralateral에도 보였으니 NEUrodegeneration causality 는 없는듯! Time Course of α-syn Inclusions and Neuroinflammation — Substantia Nigra and Agranular Insular Cortex panels (Peak Slipc Inclusions / Prolonged Toxicity / Neuronal Loss curves) |
| (Patterson, 2019 #1376) (Sortwell's lab) | aSyn PFF (recombinant but generated from mouse aSyn), Striatum (unilateral) | rats |
- 2m post injection: ipsilateral SNpc pSyn neuronal inclusion, no loss of THir neurons, and PFF perturbations of striatal TH - 4m post injection: Reduced SNpc pSyn aggregate number, ipsilateral THir neuron loss dependent perturbations of striatal TH: - 6m post injection: Minimal pSyn aggregate number in the SNpc, bilateral THir neuron loss (6m), bilateral loss of PFF dose-related striatal TH (>50%, fig 6o). (cf) Unilateral 16 μg resulted in modest sensorimotor deficits in forelimb adjusting steps associated with degeneration Flow 해석: ↑ pa (ie striatal T) neurodegeneration is a-syn being degraded into truncated ones, detectable by [antibody that recognizes pSyn,] → degeneration Ipsilateral / Contralateral panels: Peak SNpc Inclusions / Prolonged Toxicity / Neuronal Loss; series SNpc pSer pa Aggregates / SNpc Neurons / SNpc TH over Months after α-syn PFF injection |