Development of diagnostic kit and DNA aptamer for inhibiting the Fusobacterium nucleatum FadA protein reponsible for colorectal cancer

Project no. 25274 · Status: Kapanmış (Yayın bekleniyor) · 2023

Summary

<p>Colorectal&nbsp;cancer&nbsp;(CRC)&nbsp;is&nbsp;one&nbsp;of&nbsp;the&nbsp;most&nbsp;prevalent&nbsp;malignancies&nbsp;worldwide,&nbsp;and&nbsp;the&nbsp;development&nbsp;of&nbsp;specific&nbsp;biomarkers&nbsp;for&nbsp;its&nbsp;early&nbsp;diagnosis&nbsp;remains&nbsp;of&nbsp;great&nbsp;importance.&nbsp;In&nbsp;recent&nbsp;years,&nbsp;Fusobacterium&nbsp;nucleatum&nbsp;has&nbsp;been&nbsp;recognized&nbsp;as&nbsp;a&nbsp;key&nbsp;contributor&nbsp;to&nbsp;CRC&nbsp;development,&nbsp;and&nbsp;its&nbsp;adhesin&nbsp;protein,&nbsp;Fusobacterium&nbsp;adhesin&nbsp;A&nbsp;(FadA),&nbsp;has&nbsp;been&nbsp;identified&nbsp;as&nbsp;one&nbsp;of&nbsp;the&nbsp;major&nbsp;virulence&nbsp;factors&nbsp;associated&nbsp;with&nbsp;tumorigenesis.</p><p>In&nbsp;this&nbsp;project,&nbsp;we&nbsp;aimed&nbsp;to&nbsp;develop&nbsp;DNA&nbsp;aptamers&nbsp;capable&nbsp;of&nbsp;targeting&nbsp;the&nbsp;FadA&nbsp;protein&nbsp;with&nbsp;high&nbsp;specificity&nbsp;and&nbsp;affinity.&nbsp;To&nbsp;this&nbsp;end,&nbsp;recombinant&nbsp;FadA&nbsp;protein&nbsp;was&nbsp;successfully&nbsp;produced&nbsp;and&nbsp;purified,&nbsp;followed&nbsp;by&nbsp;the&nbsp;selection&nbsp;of&nbsp;target-specific&nbsp;aptamer&nbsp;candidates&nbsp;using&nbsp;the&nbsp;Systematic&nbsp;Evolution&nbsp;of&nbsp;Ligands&nbsp;by&nbsp;Exponential&nbsp;Enrichment&nbsp;(SELEX)&nbsp;method.&nbsp;The&nbsp;binding&nbsp;properties&nbsp;of&nbsp;the&nbsp;selected&nbsp;aptamers&nbsp;were&nbsp;comprehensively&nbsp;evaluated&nbsp;through&nbsp;molecular&nbsp;docking&nbsp;and&nbsp;100&nbsp;ns&nbsp;molecular&nbsp;dynamics&nbsp;simulations.</p><p>Computational&nbsp;analyses&nbsp;demonstrated&nbsp;that&nbsp;Apt1&nbsp;exhibited&nbsp;the&nbsp;most&nbsp;stable&nbsp;interactions&nbsp;with&nbsp;the&nbsp;FadA&nbsp;protein,&nbsp;characterized&nbsp;by&nbsp;lower&nbsp;binding&nbsp;free&nbsp;energy,&nbsp;a&nbsp;higher&nbsp;number&nbsp;of&nbsp;hydrogen&nbsp;bonds,&nbsp;and&nbsp;a&nbsp;more&nbsp;stable&nbsp;interaction&nbsp;profile.&nbsp;These&nbsp;findings&nbsp;suggest&nbsp;that&nbsp;the&nbsp;developed&nbsp;aptamers&nbsp;are&nbsp;promising&nbsp;candidates&nbsp;for&nbsp;use&nbsp;in&nbsp;aptamer-based&nbsp;diagnostic&nbsp;platforms&nbsp;for&nbsp;the&nbsp;detection&nbsp;of&nbsp;FadA&nbsp;and&nbsp;have&nbsp;considerable&nbsp;potential&nbsp;to&nbsp;contribute&nbsp;to&nbsp;the&nbsp;early&nbsp;diagnosis&nbsp;of&nbsp;colorectal&nbsp;cancer.</p>

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