Experimental Validation
Our team is currently focusing on the prevention of ALS, aiming to prevent stress granule formation through protein knockdown using ASOs, as well as minimizing aggregation of TDP-43 by designing an aptamer sequence that binds to the C-terminal domain of TDP-43.
Prevention of stress granule formation would effectively decrease aggregation of TDP-43 in the cytoplasm. The aptamer sequence can block TDP-43 interactions in any remaining stress granules, preventing the demixing process that contributes to ALS.
Our experiments found that ASO-A and ASO-B reduced expression of DAZAP1 by 23%, and ASO-C and ASO-D reduced expression of FAM98A by 38% and 49%, respectively. Knockdown of DAZAP1 and FAM98A significantly reduces stress granule assembly.
A limitation is that we have not yet confirmed the aptamer binds exclusively to TDP-43. We are actively working on improving staining and imaging methods to test for potential off-target interactions.
Implementation
This research has the potential to advance ALS treatment. ALS is a fatal neurodegenerative disease affecting nerves in the brain and spinal cord, leading to progressive muscle loss. There is currently no cure.
Our dual approach—using ASOs to reduce stress granule formation and an aptamer to block TDP-43 aggregation—could be developed into a therapeutic delivered via intrathecal injection into cerebrospinal fluid.
Before patient application, the aptamer sequence must be optimized, and additional tests should be performed on patient-derived ALS cells or animal models. Following preclinical validation, an Investigational New Drug (IND) application is required to begin clinical trials.
Successful trials would allow safe administration, refine dosing, and confirm therapeutic effectiveness. Ultimately, this dual-approach could limit toxic TDP-43 aggregation, enable earlier ALS detection, and improve patient outcomes.
Testing Concerns
While promising, it is crucial to ensure our research does not cause unintended effects on cells. Aptamer and ASO binding specificity must be confirmed to avoid off-target interactions with other proteins or RNA sequences.
Our findings remain preclinical. Clinical trials, safety testing, and independent validation are essential to confirm feasibility and reproducibility before any clinical application.
Ensuring reproducibility and verification by other researchers strengthens the credibility of our work and mitigates risks associated with inaccurate results.