Dr. Brian Poole, Department of Microbiology & Molecular Biology
Evaluation of how well the academic objectives of the proposal were met
We presented data from this project at two meetings, 2 presentations at the American society for Microbiology Branch Meeting, Provo UT 2010 and one at the Intermountain Research Symposium, Logan Utah 2010. At the ASM meeting one of my undergraduates was selected to give an oral presentation, and all presentations were done by students. We also published a paper with student authorship that was supported by the MEG. The paper is “Neiwold, Timothy; Clark, Daniel; Salloum, Rafeh; Poole Brian D. Interferon alpha in systemic lupus erythematosus. J Biomed Biotechnol. 2010;2010:948364” We also have three manuscripts in preparation that include data generated using MEG funding. We have generated significant and substantial data from the project and are using that data to apply for NIH funding as well as including it in manuscripts for publication.
Evaluation of the mentoring environment
I feel that the mentoring environment has been very successful. I have mentored over 22 students. Of the eight that have graduated, 5 have obtained positions doing research in industrial settings, 1 is a graduate student at BYU, 1 is working in academic research at the University of Utah and 1 is awaiting acceptance to medical school. The previously sited academic deliverables are also evidence of a productive mentoring environment. Of the 22 students I have mentored, only 2 have not continued in the lab until the present time or graduation, indicating high levels of satisfaction towards the lab. Additionally, five of my students received ORCA grants for their projects. All of my students have expressed their appreciation for the experience and have said how much they have learned.
List of students who participated and what academic deliverables they have produced or it is anticipated they will produce
| Name | Current Position | Academic Deliverables |
|---|---|---|
| Nick Whipple | PhD student at BYU | Went on to graduate School |
| Brandon Henrie | Research technician job at U of U | Author on 2 presentations |
| David Hendricks | Applying to Graduate school | |
| Brian Naylor | Applying to Dental School | |
| Doug Baumann | Research Job‐industry | uthor on 1 presentation |
| Daniel Hammond | R&D company. Applying to med school | Author on two presentations, one of which was an invited oral presentation |
| Tyler Hedman | Currently in lab | Anticipated Author on 1 manuscript , at least 2 presentations |
| Brandon Robinson | Currently in lab | Anticipated Author on 1 manuscript , at least 2 presentations |
| Kira Harris | In lab for 1 semester | |
| Kathryn Ingold | Currently in lab | Anticipated Author on 1 manuscript , at least 1 presentations |
| Trieste Bills | Currently in lab | Anticipated Author on 1 manuscript , at least 1 presentation |
| Casey Heap | Currently in lab | Anticipated Author, at least 1 presentation |
| Jason Sloan | Currently in lab | Anticipated Author on 1 manuscript , at least 1 presentation |
| Devin Burns | Currently in lab | Anticipated Author on at least 2 presentations |
| Miles Murri | Currently in lab | Anticipated Author on 1 manuscript , at least 2 presentations |
| Jared Lambert | In lab for 1 year | |
| Daniel Read | Currently in lab | Anticipated Author on 1 manuscript , at least 1 presentation |
| Sean Bergsten | Currently in lab | Anticipated Author on 1 manuscript , at least 1 presentation |
| Danny Catts | Currently in lab | Anticipated Author on 1 manuscript , at least 1 presentation |
| Amanda Stewart | Currently in lab | Anticipated Author on 1 manuscript , at least 1 presentation |
| Daniel Clark (Ph.D. Student) | Currently in lab | Author on 1 paper Author on 3 presentations Anticipated author on 3 papers |
Description of the results/findings of the project
EBV‐transformed cell lines were generated from the peripheral blood of volunteers homozygous for either the risk or protective allele at rs2004640. Nine pairs of cell lines were treated with imiquimod to induce TLR‐7 signaling. Expression of the EBV latent membrane protein 1 (LMP‐1) after treatment with imiquimod was measured using real‐time quantitative PCR. We discovered that there was a significant increase in the production of LMP‐1 after imiquimod treatment in both the risk and the protective cell lines (median 2.1‐fold, p<0.008). There was a significantly larger increase in LMP‐1 production in the cells with the risk allele compared to those with the protective allele (p=0.028) (Figure 1). Since LMP‐1 expression is upregulated by IRF7, and downregulated by IRF5 (25), the heightened production in the risk cell lines suggests that IFR5 is less effective in these cells. We confirmed that IRF7 is more active in the cells with the risk haplotype by analyzing two other genes that are primarily upregulated by IRF7 and not IRF5, calreticulin and Noxa. Both of these genes also had significantly increased expression in the risk cells after imiquimod treatment compared to the protective cells (p<0.021. 0.038, respectively) (Figure 1). There was no significant difference between risk and protective cells in upregulation of expression of IFNa‐1 or Trim22, two genes that are primarily enhanced by IRF5. [gallery link="file" columns="1" orderby="title"] Further studies on the contribution of variation at the rs2004640 locus to IRF5 splicing revealed a significant difference in the use of exon 1 variants depending on the allele present. We developed quantitative PCR primers specific for exons 1.A, 1.B, 1.C and 1.D. As we expected, intron 1.B was only present in the risk cells, since the risk allele creates this splice site. Interestingly, we observed significant reduction in the use of introns 1.C and 1.D in the cells with the risk allele, while the use of intron 1.A remained constant (Figure 2). We have also identified and sequenced 5 previously undescribed splice variants of IRF5. Three of these variants are interesting in that they use the 1.B exon, so are found exclusively in the risk cells, and have major deletions in the stability and/or protein interaction domains (Figure 3). Use of these variants in the risk cells would diminish the stability and functionality of IRF5.
Description of how the budget was spent
The budget was primarily divided between paying Daniel Clark’s graduate student stipend and laboratory supplies.
| Graduate Student Support | $6,230.79 | Supplies | $13,759 | Student Travel | $10 | Total | $20,000 |
|---|