A Bioengineering Strategy to Control ADAM10 Activity in Living Cells

Area of Science:

• Biochemistry
• Molecular Biology
• Neuroscience

Background:

• ADAM10 is a cell surface protease involved in physiological processes, with dysregulation linked to Alzheimer's disease (AD).
• Mechanisms of ADAM10 activation and its role in protein ectodomain shedding are not fully understood.
• Precise control over ADAM10 activity is needed to study its cleavage mechanisms and develop therapeutic applications.

Purpose of the Study:

• To develop a bioengineering strategy for spatiotemporal control of ADAM10 activity.
• To investigate the roles of prodomain and cytosolic tail cleavage in ADAM10 activation.
• To assess the potential of this strategy for AD gene therapy by modulating APP processing.

Main Methods:

• Engineered ADAM10 analogs with Tobacco Etch Virus protease (TEV) cleavage sites (TEVcs).
• Utilized TEV protease to induce specific cleavage of the prodomain and cytosolic tail.
• Generated ADAM10 with minimal constitutive activity, inducible by TEV or chemically activatable TEV.

Main Results:

• TEV-mediated prodomain removal alone did not activate ADAM10.
• Cleavage of the cytosolic domain significantly increased ADAM10 activity.
• Engineered ADAM10 showed increased activity upon TEV induction or chemical activation.
• The strategy promoted α-secretase activity, leading to non-amyloidogenic cleavage of APP and increased sAPPα production.

Conclusions:

• A novel bioengineering approach allows for precise, spatiotemporal control of ADAM10 activity in living cells.
• Cytosolic tail cleavage is a key regulatory step for ADAM10 proteolytic function.
• This strategy offers a potential next-generation gene therapy for AD by enhancing neuroprotective pathways.