| (Edler, 2021 #2150) | Systematic review. Overall (laura?):The team was encouraged to confirm that microglial responses in dogs are considered similar to primates (e.g. vs known differences in rodents |
| Our points | |
| NLRP3 correlates with TSPO | |
| TSPO increase in response to aSyn: ↑ in human and (AAV) RAT, But not clear in mouse (Atuka AAV aSyn mouse) |
Pipeline of Inflammation
| Azathioprine | P2, AZA-PD trial {Greenland, 2020 #1210}, ISRCTN14616801 | N=60, early PD, 12M, IMMUNE bm in blood and CSF, pet, |
Pyroptosis MOA
| [priming] | [activation] | |||
|---|---|---|---|---|
| A priming signal is provided by NF-kB-activating stimuli to transcriptionally enhance the expression of NLRP3 and pro-IL-1b (Bauernfeind et al., 2009).Many TLR and NLR ligands, as well as endogenous cytokines such as IL-1a, have been demonstrated to prime | fibrillar α-syn/ |
Aggregated aSyn activates complement to generate C5a→ ↑ C5aR1 expression IN MICROGIAL in SN TLR) (in the membrane) recognize aSyn → activation of NF-κB | Promote the production of IL-1B precursor protein (↑ gene expression) | The active caspase-1 goes out into cytosol → ↑. caspase-1 cleaves the proforms of IL-1β, IL-18 and gasdermin D (at N-term) → N terminal GSDMD oligomerize and forms a transmembrane complex (pore, inner diameter of this pore is 21.5 nm) → i) cell rupture ii) Mature IL-1β and IL-18 release from the cell through the pore (→ inflammasome 부스러기도 release 되고 주위 cell이 take up 하여 또 cycle 시작됨), IL-1β → ↑IL-6 |
| dopaminergic degeneration→ATP release→ P2X7 Receptor activation → P2X7 receptor directly interacts with NLRP3 inflammasome (Franceschini, 2015 #1789) | → [(in microglia) NLRP3-Inflammasome activation]: ↑ NLRP3 expression (= 'priming → NLR identify different stimuli Through the leucine-rich repeat (LRR) domains, → terminate its autoinhibitory state → Upon activation, NLRP3 oligomerizes through homotypic interactions between its NACHT domain to NIMA-related kinase 7 (NEK7), ] ASC binds with the altered NLR via the pyrin domain and assembles into multimers. → | |||
| intracellular debris, including cholesterol crystals (which lead to atherosclerosis), aβ particles (which are associated with AD), and | iii) ASC speck: NLRP3+ASC, 1 uM, perinuclear, punctate, → ASC (from monomer→) polymerization & filamentous→ Extracellular release of ASC→ prion-like activities of extracellular polymeric ASC can propagate chronic inflammation by spreading inflammasome signaling from cell-to-cell (20, 26). | |||
| tobacco smoke particulates (which contribute to lung disease) |
iv) inflammasome: ~0.45 uM in diameter (~700 kDa), (Kenerman 2023 AD/PD: 30nM in diameter and 50-100 nM in length ) NLRP3+ASC+caspase1→ casp1 cleavage: monomeric proenzyme (zymogen) → cleaved form (p20 or p10) → active form (dimer: p20+p10), (catalytic domain with an active site spans both the p20 and p10 subunits In some conditions, these GSDMD pores lead to lytic cell death, termed pyroptosis | |||
| Neuronal TLR2 activation → ↑AKT/Mtor→ ↓ autophagy→ ↑ a-syn→ activate complement and generate C5a → increased C5aR1 expression in primary microglia → C5a induced the secretion of IL-1 from primed microglia , | (Wang, 2016 #639) in a neuronal cell line: Caspase-1 directly cleave α-syn at Asp121 → this cleaved form was more prone to aggregation | |||
- NLRP3: NLR (NOD (neucleotide-binding oligomerization domain)-like receptor) family pyrin domain containing 3
- Pro-caspase 1: {Ball, 2020 #2006}
ASC: (apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain, inflammasome adaptor protein)
low CSHL (cold spring harbor) 2020 Abstract253 MOA
| Parkin | NEURONAL (microglial 말고) NLRP3 is a parkin substrate CSHL2020 Abstract253 | Parkin ubiquitilates NLRP3, → UPS degradation→ inflammasome priming is inhibited CSHL2020 Abstract253 | ||||
|---|---|---|---|---|---|---|
| normal | Loss of Parkin activity | ↑ accumulation of the Parkin-interacting substrate (PARIS), ZNF746 | ↑ mitoROS generation | - ↑ the priming (priming의 readout은 poster fig1!) | ||
| Parkin- | mouse |
- ↑ Neuronal NLRP3 inflammasome: {Panicker, 2022 #2067}fig1b (mouse, WB)(but =unchanged NLRP3 mRNA level) | -ASC speck (panicker 2022 #2067}fib1d (mouse, IHC) (but not in microglia, fig1d) | |||
| -casp1 p20 (ie ↑ increased cleavage) 2022 #2067}fib1b (mouse, WB) (but not in microglia, fig1c, IHC) (but =unchanged CASP1 mRNA level) | ↑ DA neuron death (Panicker, 2020 #1815) | |||||
| In vitro | {Panicker, 2022 #2067}-And in PARKIN-deficient hDA neurons | {Panicker, 2022 #2067} CSHL2020 Abstract253 | ||||
| {Panicker, 2022 #2067}-And in Prkn-deficient hDA neurons | {Panicker, 2022 #2067}-And in PARKIN-deficient hDA neurons | |||||
| a lack of induction of A20 (A20 is a microglia-enriched protein and a well-known negative regulator of the NF-κB pathway) {Mouton-Liger, 2018 #1216} | ↑ Microglial NLRP3 inflammasome activation {Mouton-Liger, 2018 #1216, KO Mice-derived microglial culture) | (Mouton-Liger, 2018 #1216) ↑ IL-1b and IL-18 in Park22/2 and Pink12/2 cells. This defect was confirmed in blood-derived macrophages from patients with PARK2 mutations and was reversed by MCC950 | ||||
| =parkin protein, but ↓ parkin autoubiquitination (wb, 2022 panicker #20670 | ||||||
| - ↑ NLRP3 (2022 Panicker #2067}fib6b (wb) | - ↑ casp1 p20 (2022 #2067}fib6b (wb) | ↑ DA neuron death (2022 #2067}fib6 (wb) |
ptosis evidence/MOA/Correction in PD
| Gene | Microglia activation | Inflammasome | Downstream (cytokine) | Cell death ) | DA (& metabolites) | aSyn | ||
|---|---|---|---|---|---|---|---|---|
| Brain (Postmortem) | ↑ CD4+ T lymphocytes infiltration (SN & amygdala), increased inflammatory markers such as | (Gordon, 2018 #585) fig 1) IBA1+ cells (SN):staining figure but no quantification | (Gordon, 2018 #585,n=6 each, PD stage not specified) fig 1) ↑ NLRP3 & ASC (IF) in IBA-1-positive microglia in SN of PD patient (정량화x), 아래그림은(fig1b) WB. |
- (Mogi, 1994 #654) PD (n=9), PD stage not specified, : In striatum & Frontal cortex in PDD/PDND) above table mean w SEM ↑ IL-1β , ↑ TLR4 내가 sd로 바꿔 d 계산하니 1.03 for IL-1b | Not mention | (Wang, 2016 #639)Costaining of αSyn and caspase-1 in the autopsy brain of PD patients. |
Uncertain Spans
| location | transcription | uncertainty |
|---|---|---|
| Pro-caspase 1 ribbon diagram (Ball, 2020 #2006) | residue labels and kDa annotations | The diagram is preserved as evidence; individual residue tag positions on the ribbon are too small for cross-validation against the photo. |
| Pyroptosis evidence/MOA Brain row inset bar chart | y-axis labels “Fold change” and bar values | Inline bar chart is kept as evidence; bar heights are not transcribed. |
| Mogi 1994 inline table | numeric SEM values and (%) percentages | Several digits in the four-row IL-1β / IL-6 table are partially hidden by yellow highlight overlays. |
| CSHL 2020 Abstract253 MOA table | row “=parkin protein, but ↓ parkin autoubiquitination (wb, 2022 panicker #20670” | Closing brace of the citation appears written as “0” rather than ”}”; preserved as seen. |
| Page top continuation row | ”Our points” then blank cell | Cell to the right of “Our points” is blank in the photo; cannot confirm whether content is on a continuation page. |