Protein assembly modulation: A new approach to Amyotrophic Lateral Sclerosis (ALS) therapeutics

Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease with a complex, multifactorial pathophysiology, most commonly manifest as loss of motor neurons. We introduce a new mechanism of ALS pathogenesis via a novel drug-like small molecule series that targets a subset of protein disulfide isomerase (PDI) within a previously unappreciated transient and energy-dependent multi-protein complex. This drug was found to have activity in cellular models for both familial and sporadic ALS, as well as in transgenic worms, flies, and mice bearing a diversity of human genes with ALS-associated mutations. The hit compound was initially identified as a modulator of human immunodeficiency virus (HIV) capsid assembly in cell-free protein synthesis and assembly (CFPSA) systems, with demonstrated antiviral activity in cell culture. Its advancement for ALS-therapeutics, and subsequent separation of activity against HIV and ALS into separate chemical subseries through structure-activity-relations..., In vitro studies
Stress granule aggregation in SH-SY5Y cells overexpressing TDP-43
A TDP-43 cellular model was successfully generated by establishing stable cell-lines over-expressing wild-type TDP-43 and M337V mutant TDP-43 and showing the display of stress granules, following arsenite treatment. SH-SY5Y tet-on TDP-43 partial 3`-UTR (wt or M337V), 50.000 cells (DMEM/F12 + 1 µg/mL doxycycline) incubated on cover slips overnight. Cells were treated sodium arsenite at a final concentration of 250 µM and incubated for 90 min. Cells were washed with 500 µL PBS.  Cells were fixed with 4% PFA in PBS pH 7.4 for 15 min at RT.  Cells were washed with 500 µL PBS. Permeabilize/ block cells with 5% milk powder, 1% BSA and 0.5 % saponin in PBS for 45 min @ RT. Add respective {TDP-43 C-terminal domain Antibody (1:1000; Purchased from Proteintech), HuR (1:500; Purchased from Santa Cruz} in 1% PBS and 0.5% saponine in PBS and incubated overnight at  4°C.  Cells were washed 3 x with 500 µL PBS and incub..., , # Protein Assembly Modulation: A New Approach to Amyotrophic Lateral Sclerosis (ALS) Therapeutics

[https://doi.org/10.5061/dryad.fqz612k0k](https://doi.org/10.5061/dryad.fqz612k0k)

**Overview:**

The data for this paper is available in two files: one PowerPoint and one Excel file. The PowerPoint contains scans of western blots and the excel file contains the raw data used in the paper.
 
**Power point file description:**

Each slide in the PowerPoint contains scans of the western blots that were quantified and plotted in figure 5. Each lane of the western blot is labelled numerically at the top of the gel and a key is provided on the right side of the gel, detailing what sample is present in each lane.   

**Excel file description:**

Each tab in the Excel file contains the raw data corresponding to a specific figure. Tabs are labeled according to the figure number, with supplemental figures indicated by an 's' before the number.

In this Excel file, cells containing \"n/a\" represent m...