Specific Aim 2: Characterize mucosal MmuPV1 infection and persistence in nude mice by RNA sequencing analysis.
Rationale: Both outbred and inbred nude mice are susceptible to MmuPV1 infection at mucosal sites including tongue, vagina and anus (16-18). Although all three sites are susceptible to MmuPV1 infection, the disease outcome varies. Vaginal tract are the most susceptible sites among the three and the infection can be detected along the vaginal tract and up to the cervical region (16, 17). For oral infection, the infection was prevalent at the single circumvallate papilla of the mouse tongue and the adjacent Von Ebners glands at the dorsal tongue (18). For anal infection, the infection can be detected mostly at the transformation zone …show more content…
Although the secondary infections usually occurred at a later time point, it might happen in our proposed studies. If we detect cutaneous lesions in our animals before we terminate the experiment, we will harvest these lesions for further investigation. These lesions will be proper samples to compare the transcripts between the cutaneous and the mucosal sites of the same animals. Dr. Yuka Imamura Kawasawa and her team will conduct the RNA sequencing and downstream data analyses. For the vaginal tract, we plan to divide it into lower, middle and upper sections to examine whether any difference exists in these different regions. In addition, the cervical region can be analyzed separately. We expect to see significant differences in signaling pathways between the infected vs non-infected animals based on previous studies in human and animals (our preliminary data)(49). Molecules with significantly higher or lower levels will be further confirmed with microarrays and real-time RT-PCR analysis. We will also define these molecules at the protein level using immunohistochemistry or western blot analysis. Many factors such as hormonal influences, hemoglobin contamination, pH, and cervical secretion volume all contribute to the inherent variation of local levels (50, 51). We will take these factors into consideration in our sample collections. We will expand our mucosal study to male mice after we collect the data. The oral and anal infections have showed gender biases in the human population (). It would be interesting to find out this is true in our mouse model. The current study will enable us to define key candidates for potential therapeutic strategies against papillomavirus-associated
Rationale: Both outbred and inbred nude mice are susceptible to MmuPV1 infection at mucosal sites including tongue, vagina and anus (16-18). Although all three sites are susceptible to MmuPV1 infection, the disease outcome varies. Vaginal tract are the most susceptible sites among the three and the infection can be detected along the vaginal tract and up to the cervical region (16, 17). For oral infection, the infection was prevalent at the single circumvallate papilla of the mouse tongue and the adjacent Von Ebners glands at the dorsal tongue (18). For anal infection, the infection can be detected mostly at the transformation zone …show more content…
Although the secondary infections usually occurred at a later time point, it might happen in our proposed studies. If we detect cutaneous lesions in our animals before we terminate the experiment, we will harvest these lesions for further investigation. These lesions will be proper samples to compare the transcripts between the cutaneous and the mucosal sites of the same animals. Dr. Yuka Imamura Kawasawa and her team will conduct the RNA sequencing and downstream data analyses. For the vaginal tract, we plan to divide it into lower, middle and upper sections to examine whether any difference exists in these different regions. In addition, the cervical region can be analyzed separately. We expect to see significant differences in signaling pathways between the infected vs non-infected animals based on previous studies in human and animals (our preliminary data)(49). Molecules with significantly higher or lower levels will be further confirmed with microarrays and real-time RT-PCR analysis. We will also define these molecules at the protein level using immunohistochemistry or western blot analysis. Many factors such as hormonal influences, hemoglobin contamination, pH, and cervical secretion volume all contribute to the inherent variation of local levels (50, 51). We will take these factors into consideration in our sample collections. We will expand our mucosal study to male mice after we collect the data. The oral and anal infections have showed gender biases in the human population (). It would be interesting to find out this is true in our mouse model. The current study will enable us to define key candidates for potential therapeutic strategies against papillomavirus-associated