Correction studies tail, Message 모음 (GBA activity / GlcCer / GlcSph / a-syn), Goals, w reduced GBA activity Gene Therapy
Correction studies (continued)
| model | phenotype | intervention | outcome | label | |
|---|---|---|---|---|---|
| A53T α-syn Mouse, 4 month old | ↓ GBA activity, ↑ a-syn, GlcCer/GlcSph 는 언급도 없음. | AAV striatum | Fig 4. GBA activity 10배증가 → aSyn 잘 봐주면 반 감소. Proteinase K-resistant α-Synuclein Immunoreactivity panel labels: Surgery at 4 months, Surgery at 12 months, AAV. | PD only model | |
| (Schondorf et al. 2014, PMID 24905578) | iPSC-derived dopaminergic neurons from GBA-PD patients | ↑ a-syn, ↑ GlcCer, ↓ GBA activity, ↑ GBA protein, Fig8) ↑ LAMP1, ↑ LC3II (basal), ↓ LC3 influx, altered calcium and neuronal vulnerability | Gene correction | ↑ aSyn in this iPS DA neurons from GBA-PD patients asyn, GBA activity, calcium and neuronal vulnerability, 이것들이 다 correction되었다는 건지 잘 몰겠네. Fig8) ↑ LAMP1, ↑ LC3II (basal), ↓ LC3 influx, (these all corrected) | GBA-PD model |
| (Bae, 2014 #610) | in vitro V1S cells | aSyn spread | Reduced cell-to-cell transmission of aSyn in V1S cells (when the cells were cocultured with SV2GBA -/- cells) was reversed by AAV vector-mediated ectopic expression of the wild-type GBA1 | ||
| (Magalhaes, 2016 #746) | In vitro Gba1 heteroz mouse embryonic fibroblasts (MEF) | lysosome | Fig2 D) Cerezyme (exogenous GBA treatment) Increased the acidic vesicles (lysosome) |
Message 모음 (GBA activity-GlcCer GlcSph - a-syn)
아래 중 노란것들이 GBA boosting 에 relevant
SNCA/SNCA
| mice | GBA activity | GlcCer | GlcSph | p-a-syn |
|---|---|---|---|---|
| SNCA/SNCA | ||||
| Dose | ||||
| Tx effect Ambroxol | ↑ (<10%) | Not done | Not done | ↓ (40%) |
| message |
CBE mice / PR001
| mice | GBA activity | GlcCer/GlcSph | a-syn |
|---|---|---|---|
| CBE mice / PR001 | cortex 3.2×1010 ↑ normal control보다 훨씬 더 증가시킴 | Cortex 3.2×1010 ↓ normal control에 근접 | na |
| message | GBA activity를 normal보다 훨씬 더 올려야 GlcCer/GlcSph는 normalize 됨. |
4L/PS-NA mice / PR001
| mice | GBA activity | GlcCer/GlcSph | a-syn |
|---|---|---|---|
| 4L/PS-NA mice / PR001 | cortex 4.3×1010 (dose 2) ↑ 15× | cerebellum 4.3×1010 (dose 2) = | Cortex 2.4×1010 ↓ 60% |
| message | GlcCer/GlcSph 변화 없이도 GBA activity 변화만으로 a-syn 변화시킴. |
Venglu / s-181
| mice | GBA activity | GlcCer | GlcSph | a-syn |
|---|---|---|---|---|
| GbaD409V/D409V | Whole brain | Cortex | Cortex | a-syn (hippo, insoluble) na |
| Tx effect Venglu | No effect (GCSi니까 당연) | ↓ (20%) | ↓ (50%) | ↓ to baseline (not revert) |
| CBE / Tx effect Venglu | (GCSi니까 무의미) | ↓ (>50%) | ↓ (50%) | na |
| A53T-SNCA / Tx effect Venglu | (GCSi니까 무의미) | ↓ (30%) | NA / No accumulation | ↓ (20%) |
| GbaD409V/+ / Tx effect s-181 | 아마 whole brain ↑ (15%) | ↓ (10%) | ↓ (60%) | a-syn (insoluble) ↓ (50%) |
| message | GlcSph는 downstream 이니까 change 증폭, & a-syn 도 GlcSph 따라감? |
[summary of messages]
A-syn more correlates with GlcSph than with GlcCer.
GlcSph is downstream and more sensitive and change (그래서 더 크게 움직이긴 하는데), but normalize 시키려면 GBA Activity 를 normal control보다 훨씬 더 올려야 함.
Venglu는 PD P2에서 CSF GlcSph 검출 못 해서 감소 못 보여줬는데, brain 상황은 불명 (brain에서도 축적 없었는지, or csf와 correlation 이 없는 건지), GD P2에서는 ↓ CSF GlcSph (by 48%), ↓ CSF GlcSph (by 75%)
| (Charkhand et al. 2019, PMID 31467847), patient 1 (12y) | ambroxol (3 m) → ↓ CSF GlcSph (by 60%) → ataxia improved, but seizure and EEG remained unchanged despite ambroxol 3 y. |
| Patient 2 (21y) | ambroxol (6 m) → ↓ blood GlcSph (by 60%) → ataxia improved, seizure의 frequency는 그대로나 duration이 줄어 status 안 발생. |
Goals
| domain | message |
|---|---|
| GD | GBA activity를 normal 보다 훨씬 더 올려야 (혹은 normal 수준으로) GlcCer/GlcSph 는 normalize 됨 (GBA activity 의 moderate 증가 (ie still below normal control)로는 GlcSph 도 moderate 감소 밖에 안 됨) |
| GD | Goal: GlcSph < 25% (이유: D GlcSph accumulation 이 원래 많고, gene dose effect 생각하고, 소아 case 사례 생각하면) |
| PD | ↑ GBA activity by 10% (ambroxol), 15% (s-181) → [↓?] a-syn (half) |
| PD | GlcSph reduction by 50% → [↓?] a-syn (50%) (s-181) (venglu), Goal: ↓ A-syn 50% |
w reduced GBA activity Gene Therapy
| PS | PK | TE / Target occupancy | TF (Target Function) | Pathway Modulation | Disease Modifying | Physiologic response | Pathophysiologic process | Clinical endpoint | ||
|---|---|---|---|---|---|---|---|---|---|---|
| Genotype/phenotype |
1. GBA mutation 2. reduced GCase activity without mutations ↓ GBA in CSF (?) & lymphocytes/blood | GBA protein level, | GBA activity (ambroxol CSF잼) GCase in lymphocytes, GBA in blood (Alcalay 2015) | Glucosylceramide Glucosylsphingosine (Gegg 2015 에서 brain 에서 안 늘었으니) Glucosylceramide 는 무의미하겠네 | Autophagy? calcium? ER stress? | a-syn CSF) biomarkers levels of a-syn | ↑ markers of macrophage activation: angiotensin-converting enzyme, cathepsin S, chitotriosidase, and CCL18 in the blood plasma; and TNFα in splenic Gaucher cells | |||
| clinical selection note |
(cf glucosylceramide & glucosylsphingosine are not increased in PD, PD+GBA postmortem Gegg2015) Confirmed dopamine deficit and Exclude SWEDD (by DATScan) ((Stephenson et al. 2019, PMID 31306141) | |||||||||
| Allometric scaling | GBA protein level (Ambroxol → ↑ GBA levels in non-human primates [28].) | GBA activity (oral Ambroxol → for 12 days ↑ GBA activity in the brainstem, midbrain, cortex, and striatum of wild-type as well as transgenic mice carrying a L444P Mutation) | ↓ substrate (in neuron?) | α-syn → Ambroxol had a reduction in α-syn levels in the brainstem and striatum of transgenic mice overexpressing human α-syn | α-syn S129 → Ambroxol reduced S129 phosphorylation of α-syn (suggested to play a critical role in α-syn aggregation and formation of Lewy bodies and neuritis) in the brainstem by 41% | ABX treatment significantly increased N370S and F213I mutant GCase activity and protein levels in GD fibroblasts → enhancement of GCase activity and a reduction in glucosylceramide storage was verified in ABX-treated GD lymphoblasts (N370S/N370S). | ||||
Uncertain Spans
| location | transcription | uncertainty |
|---|---|---|
| Allometric / ABX row | N370S and F213I mutant GCase activity | reads as written; the F213I may also read as F2131 in OCR, preserved verbatim. |