CDA email thread tail, CEI / Companies CEI / Vesalius distinct clinical feature types, Ambagon Argo molecular-glue 14-3-3 client table, Brain / CSF detection table, Giusto 2021 14-3-3 review, Underwood 2021 PFF mice notes

CDA email thread (continued)

	Best Jaewon	Thank you and Cheers! Lisa Cabeceiras Best Jaewon
		α

CEI

Companies CEI

Vesalius	DISTINCT CLINICAL FEATURE TYPES ™ Candidate circuits (w signaling parter) Dependency target target Eg motor+sleep (then what endpoint?), but p5 fig: y-axis has only UPDRS III? Rapid progression? How are they distinct? How to identify/screen them (eg gene-blood cyctokine-brain cytokine - clinical feature?) How to connect these two? Eg. inflammation Like patway analysis? - Circuit A: inflammation - Circuit B: neurodegeneration validation ??????? validated in native genome models validated in native genome models -
Ambagon Argo	a molecular glue approach, not to degrade proteins directly but instead to modulate functions of 14-3-3, a scientific seminar from the Ambagon's CTO, Christian Ottmann https://www.youtube.com/watch?v=0LZ4sNhKbmc
	Client protein: Mostly phosphorylated proteins Desired consequence aSyn i) stabilizing the naturally occurring 14-3-3:aSyn interaction ii) (eventually) Degrade In human LBs, 14-3-3 is found, 14-3-3 binds to LMW multimers of aS → 'traps' these multimers and prevents formation of HMWaS multimers (These molecules interact with a binding groove (정확히 어디? The 14-3-3 encyclopedia provides a systematic understanding of the 14-3-3:client interface) that is formed when 14-3-3 recognizes phosphorylated serine- or threonine-containing motifs on α-syn) → ↓ aggregation (oligomerization) → ↓ spread → degraded in aggresome (note: aggresome refers to an aggregation of misfolded proteins in the cell, formed when the protein degradation system of the cell is overwhelmed. An aggresome forms around the microtubule organizing center in eukaryotic cells, adjacent to or enveloping the cell's centrosomes. The aggresome is eventually targeted for autophagic clearance) LRRK2 14-3-3 bind missensed LRRK2? / p-LRRK2 → Stabilize? De-activate? Parkin GBA : p- 아니라서 대상 안 되겠군. 14-3-3 bind missensed GBA → Stabilize → escape ER degradation

Seven genes encode seven distinct 14-3-3 proteins

There are common recognition motifs for 14-3-3 proteins that contain a phosphorylated serine or threonine residue

• molecular glue
  • is a small molecule
  • Distinct from PROTAC molecules, molecular glues insert into a naturally occurring PPI interface, with contacts optimized for both the substrate and ligase within the same small molecule entity.[4][5]
• Questions
  • Selectivity (off target)
  • 14-3-3 expression level in PD
  • LRRK2
    • Safety has been an issue
      • Just normalization (return to the baseline) would be ok?

	Brain	CSF	Detection rate
HC	Kawamoto 2002: In the normal and neurodegenerative conditions, 14-3-3-like immunoreactivity was observed mainly in the neuronal somata and proximal processes.	{Zerr, 1998 #2252} Rarely detected
Non-CJD		{Satoh 2010} Meta-analysis 에 안 잡힘
PD	[Kawamoto 2002: strong 14-3-3-like immunoreactivity in both classical and cortical Lewy bodies in the brains affected by PD and DLBD. both classical and cortical Lewy bodies were intensely immunolabeled and some dystrophic neurites were also immunoreactive for 14-3-3. Our results suggest that 14-3-3 proteins may be associated with Lewy body formation	잡힘 {Xiang 2022 #2254}	~30%

Reading: ]Giusto 2021 #2246] REVIEW ARTICLE OPEN ‘Pathways to Parkinson’s disease: a spotlight on 14-3-3 proteins’

Two primary binding sites:

• pS910 (14-3-3γ EC50 = 2.5 μM, 14-3-3σ (이건 sigma 인데, 혹시 δ delta 의 오기?) EC50 = 27.7 μM)
• pS935 (14-3-3γ EC50 = 19.8 μM, 14-3-3σ EC50 = 183.7 μM)

Di Maio et al., 2018: in PD: postmortem SN section (fig 2): ↑LRRK2 activity, ↑4-fold phosphorylation (pThr73) of the LRRK2 substrate Rab10, ↓LRRK2-14-3-3 binding.

The increase in LRRK2 activation state in iPD dopamine neurons corresponded to ↓5x in the 14-3-3 PL signal

PD DATA: Human, cell, animal

• In a “dopaminergic” neuroblastoma cell line investigators were able to demonstrate that stable overexpression of certain 14-3-3 isoforms were protective against rotenone and MPTP induced toxicity and utilized a C. elegans model of PD to show that human 14-3-3 and its homolog (ftt2) protect against α-syn toxicity. (Yacoubian TA et al., 2010)
• Plotegher et al. 2014 utilized a number of biochemical approaches to show that a particular isoform of 14-3-3 interacts specifically with α-syn oligomers (not monomers and not preformed fibrils (PFFs)) and 14-3-3 inhibited α-syn toxicity in an engineered HEK cell system overexpressing α-syn (Plotegher et al., 2014)
• Using a mouse α-syn PFF model investigators found that overexpression of 14-3-3 delayed α-syn inclusion formation whereas the 14-3-3 inhibitor peptide difopein accelerated aggregate formation and showed reduced TH+ neuronal counts (Underwood R et al., 2021)
• Wang et al. 2018 demonstrated that overexpression of 14-3-3 reduced oligomeric α-syn, prevented formation of pathological insoluble α-syn after PFF treatment in vitro and reduced toxicity of released α-syn in an in vitro paracrine model. Conversely, the 14-3-3 inhibitor peptide difopein increased α-syn toxicity in mouse primary hippocampal cells as well as differentiated human SH-SY5Y neuronal cells.
• The tool molecule Fusicoccin-A (Fc-A), that stabilizes 14-3-3 α-syn interactions, produced a moderate but significant protection of dopaminergic neurons in in vitro rat and in vivo mouse models of PD (Vinueza-Gavilanes et al., 2023).

{Underwood, 2021 #2713}

• in PFF mice: 14-3-3 θ transgene overexpression in cortical and amygdala regions rescued social dominance deficits induced by PFFs at 6 months post injection, whereas 14-3-3 inhibition by transgene expression of the competitive 14-3-3 peptide inhibitor difopein in the cortex and amygdala accelerated social dominance deficits. The behavioral rescue by 14-3-3 θ overexpression was associated with delayed α syn aggregation induced by PFFs in these brain regions. Conversely, 14-3-3 inhibition by difopein in the cortex and amygdala accelerated α syn aggregation and reduction in NECAB1-positive neuron counts induced by PFFs.
• 14-3-3 θ overexpression by AAV in SN also delayed α syn aggregation in the SN and partially rescued PFF-induced reduction in TH-positive dopaminergic cells in the SN. 14-3-3 inhibition in the SN accelerated nigral α syn aggregation and enhanced PFF-induced reduction in TH-positive dopaminergic cells

Uncertain Spans

location	transcription	uncertainty
CDA tail row 2	α	a single Greek alpha character sits in the second cell of the trailing row; possibly an alpha-syn placeholder or a stray symbol; preserved verbatim.
aSyn small mini-table	peptide cells TVEGAGAxIAAA, GSIAAAIGFVK, EAYEMPxEEGY	OCR is partial through small font; preserved verbatim with x for illegible character positions.