Abstract Title
Determining the Benefit Human Cytomegalovirus Gains by Down Regulating Nidogen-1
Acknowledgement of Funding Sources
Other
Additional Funding Sources
This work was supported by NIH grants RO1 AIO51563, INBRE program P20 GM103408 and COBRE program P20 RR015587. The project described was support through the University of Idaho Summer Undergraduate Research Fellowship made possible by a 2017-2018 Undergraduate Research Grant from the Higher Education Research Council/Idaho State Board of Education.
Abstract
Human Cytomegalovirus (HCMV), is known to be the most prevalent cause of neurological birth defects, ranging from microcephaly to sensorineural hearing loss. This study aims to elucidate the benefit HCMV derives from modifying a particular cellular mechanism to more efficiently disperse infected cells. Within 6-8 h post infection, HCMV begins to downregulate Nidogen-1 (NID1), an important component of the extracellular matrix (ECM) secreted by endothelial cells, by both protein stability and decreased mRNA transcription. To determine if the absence of NID1 increases dispersal of HCMV, we designed a series of transmigration assays that utilize human umbilical vein endothelial cells (HUVECs) seeded onto a polycarbonate membrane. THP-1 monocytes were then seeded on top of the HUVEC monolayer and total transmigration of the monocytes was measured after 24 and 48 hours. Five different treatments of HUVECs, ranging from full HCMV infection to NID1 knockdown, were used to test our hypothesis that downregulation of NID1 increases transmigration due to decreased ECM integrity. Preliminary results with uninfected monolayers yielded an average of 24% transmigration vs NID1 knockdown at an average of 35% transmigration. Ultimately, we believe the targeting of NID1 provides HCMV a selective advantage, which exacts a negative toll on the developing fetus.
Determining the Benefit Human Cytomegalovirus Gains by Down Regulating Nidogen-1
Human Cytomegalovirus (HCMV), is known to be the most prevalent cause of neurological birth defects, ranging from microcephaly to sensorineural hearing loss. This study aims to elucidate the benefit HCMV derives from modifying a particular cellular mechanism to more efficiently disperse infected cells. Within 6-8 h post infection, HCMV begins to downregulate Nidogen-1 (NID1), an important component of the extracellular matrix (ECM) secreted by endothelial cells, by both protein stability and decreased mRNA transcription. To determine if the absence of NID1 increases dispersal of HCMV, we designed a series of transmigration assays that utilize human umbilical vein endothelial cells (HUVECs) seeded onto a polycarbonate membrane. THP-1 monocytes were then seeded on top of the HUVEC monolayer and total transmigration of the monocytes was measured after 24 and 48 hours. Five different treatments of HUVECs, ranging from full HCMV infection to NID1 knockdown, were used to test our hypothesis that downregulation of NID1 increases transmigration due to decreased ECM integrity. Preliminary results with uninfected monolayers yielded an average of 24% transmigration vs NID1 knockdown at an average of 35% transmigration. Ultimately, we believe the targeting of NID1 provides HCMV a selective advantage, which exacts a negative toll on the developing fetus.