Characterizing Neonatal Human Epidermal Keratinocytes-derived Exosomes
Session Number
MEDH 17
Advisor(s)
James Coy-Dibley (PhD Candidate)
Daniela M Menichella (PI), Northwestern University
Discipline
Medical and Health Sciences
Start Date
17-4-2024 10:45 AM
End Date
17-4-2024 11:00 AM
Abstract
Painful diabetic neuropathy (PDN) is a debilitating complication of diabetes with patients suffering from a painful, burning sensation in their extremities. Available treatments have limited effect in masking the pain without addressing the underlying mechanisms of the disease, which are currently poorly understood. Keratinocytes are in close proximity to cutaneous nerve terminals, enabling bidirectional communication between keratinocytes and cutaneous nerves. One mode of communication that is understudied in the skin is extracellular vesicles (EVs), namely exosomes, which are secrete nanoparticles of size range 30-150nm. To study keratinocyte-derived exosomes, we employed the immortalized neonatal human epidermal keratinocyte (NHEK) cell line and isolated NHEK-derived exosomes via size exclusion chromatography. We morphologically characterized the NHEK nanoparticles via dynamic light scattering and electron microscopy before molecularly confirming the exosomes with western blotting for known molecular markers. We confirmed the functional uptake of the isolated exosomes with DIR-labeling. These findings will be translated into primary cultures and in-vivo functional studies in the pursuit of uncovering novel treatments for PDN.
Characterizing Neonatal Human Epidermal Keratinocytes-derived Exosomes
Painful diabetic neuropathy (PDN) is a debilitating complication of diabetes with patients suffering from a painful, burning sensation in their extremities. Available treatments have limited effect in masking the pain without addressing the underlying mechanisms of the disease, which are currently poorly understood. Keratinocytes are in close proximity to cutaneous nerve terminals, enabling bidirectional communication between keratinocytes and cutaneous nerves. One mode of communication that is understudied in the skin is extracellular vesicles (EVs), namely exosomes, which are secrete nanoparticles of size range 30-150nm. To study keratinocyte-derived exosomes, we employed the immortalized neonatal human epidermal keratinocyte (NHEK) cell line and isolated NHEK-derived exosomes via size exclusion chromatography. We morphologically characterized the NHEK nanoparticles via dynamic light scattering and electron microscopy before molecularly confirming the exosomes with western blotting for known molecular markers. We confirmed the functional uptake of the isolated exosomes with DIR-labeling. These findings will be translated into primary cultures and in-vivo functional studies in the pursuit of uncovering novel treatments for PDN.