Investigation of the Potential Use of Stem Cell-Derived Exosomes as Drug Carriers in Neurodegenerative Diseases

Project no. 32232 · Status: Kapanmış (Yayın bekleniyor) · 2024

Summary

<p class="ql-align-justify"><span style="background-color: transparent; color: rgb(0, 0, 0);">This&nbsp;study&nbsp;aimed&nbsp;to&nbsp;create&nbsp;an&nbsp;in&nbsp;vitro&nbsp;model&nbsp;of&nbsp;Parkinson's&nbsp;Disease&nbsp;(PD).&nbsp;Because&nbsp;experimental&nbsp;animals&nbsp;cannot&nbsp;fully&nbsp;replicate&nbsp;human&nbsp;physiology,&nbsp;the&nbsp;goal&nbsp;was&nbsp;to&nbsp;create&nbsp;a&nbsp;3D&nbsp;microenvironment&nbsp;using&nbsp;human&nbsp;cells,&nbsp;closer&nbsp;to&nbsp;in&nbsp;vivo&nbsp;conditions,&nbsp;and&nbsp;to&nbsp;apply&nbsp;this&nbsp;model&nbsp;in&nbsp;a&nbsp;3D&nbsp;PD&nbsp;model&nbsp;by&nbsp;encapsulating&nbsp;exosomes&nbsp;derived&nbsp;from&nbsp;stem&nbsp;cells&nbsp;with&nbsp;levodopa&nbsp;(L-dopa),&nbsp;commonly&nbsp;used&nbsp;in&nbsp;PD,&nbsp;under&nbsp;Exo-Ld&nbsp;conditions.&nbsp;Since&nbsp;the&nbsp;effectiveness&nbsp;of&nbsp;commonly&nbsp;used&nbsp;drugs&nbsp;in&nbsp;PD&nbsp;diminishes&nbsp;before&nbsp;reaching&nbsp;the&nbsp;brain,&nbsp;they&nbsp;are&nbsp;used&nbsp;at&nbsp;increased&nbsp;doses.&nbsp;Because&nbsp;these&nbsp;high&nbsp;doses&nbsp;have&nbsp;side&nbsp;effects,&nbsp;the&nbsp;effectiveness&nbsp;of&nbsp;encapsulating&nbsp;L-dopa&nbsp;with&nbsp;exosomes&nbsp;(EVs),&nbsp;which&nbsp;act&nbsp;as&nbsp;cargo&nbsp;in&nbsp;the&nbsp;human&nbsp;body,&nbsp;to&nbsp;increase&nbsp;its&nbsp;effectiveness&nbsp;and&nbsp;facilitate&nbsp;targeted&nbsp;therapy&nbsp;was&nbsp;investigated.&nbsp;In&nbsp;this&nbsp;thesis,&nbsp;EV&nbsp;characterization&nbsp;was&nbsp;performed&nbsp;using&nbsp;Raman&nbsp;Spectroscopy,&nbsp;considering&nbsp;the&nbsp;lengthy&nbsp;and&nbsp;challenging&nbsp;optimizations&nbsp;of&nbsp;existing&nbsp;approaches.&nbsp;L-dopa&nbsp;will&nbsp;be&nbsp;encapsulated&nbsp;into&nbsp;exosomes&nbsp;obtained&nbsp;from&nbsp;stem&nbsp;cells&nbsp;(SSCs)&nbsp;using&nbsp;a&nbsp;sonication&nbsp;device.&nbsp;In&nbsp;a&nbsp;3D&nbsp;</span><em style="background-color: transparent; color: rgb(0, 0, 0);">in&nbsp;vitro</em><span style="background-color: transparent; color: rgb(0, 0, 0);">&nbsp;PH&nbsp;model,&nbsp;the&nbsp;effect&nbsp;of&nbsp;L-dopa&nbsp;encapsulated&nbsp;in&nbsp;exosomes&nbsp;on&nbsp;dopaminergic&nbsp;neuron&nbsp;regeneration&nbsp;was&nbsp;evaluated&nbsp;molecularly&nbsp;by&nbsp;qPCR&nbsp;analysis,&nbsp;and&nbsp;specific&nbsp;antibodies&nbsp;were&nbsp;evaluated&nbsp;by&nbsp;immunofluorescence&nbsp;analysis.&nbsp;It&nbsp;was&nbsp;proven&nbsp;by&nbsp;qPCR&nbsp;and&nbsp;immunofluorescence&nbsp;analyses&nbsp;that&nbsp;6-OHDA&nbsp;application&nbsp;increased&nbsp;α-synuclein&nbsp;(α-syn)&nbsp;expression.&nbsp;Furthermore,&nbsp;it&nbsp;was&nbsp;shown&nbsp;that&nbsp;6-OHDA&nbsp;application&nbsp;decreased&nbsp;tyrosine&nbsp;hydroxylase&nbsp;(TH)&nbsp;and&nbsp;vasointestinal&nbsp;peptide&nbsp;(VIP)&nbsp;immunofluorescence&nbsp;expression,&nbsp;and&nbsp;showed&nbsp;a&nbsp;reducing&nbsp;effect&nbsp;on&nbsp;TH&nbsp;and&nbsp;dopamine&nbsp;transporter&nbsp;Slc6a3&nbsp;gene&nbsp;expression.&nbsp;It&nbsp;was&nbsp;semi-quantitatively&nbsp;proven&nbsp;by&nbsp;immunofluorescence&nbsp;analysis&nbsp;that&nbsp;EV,&nbsp;L-dopa,&nbsp;and&nbsp;L-dopa-loaded&nbsp;EV&nbsp;treatment&nbsp;decreased&nbsp;α-syn&nbsp;expression&nbsp;and&nbsp;increased&nbsp;TH&nbsp;and&nbsp;VIP&nbsp;expression.&nbsp;EV&nbsp;and&nbsp;L-dopa-loaded&nbsp;EV&nbsp;treatment&nbsp;demonstrated&nbsp;a&nbsp;regulatory&nbsp;effect&nbsp;on&nbsp;TH&nbsp;and&nbsp;Slc6a3&nbsp;gene&nbsp;expression.</span></p><p></p>

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