Aaron Mcclelland Email & Phone Number

Seattle, Washington, United States

Seattle, Washington, United States

Studied the effects of aging and also focal segmental glomerulosclerosis on kidney health. Work was specifically focused on granular juxtaglomerular cells and their ability to migrate to sites of podocyte loss where these cells undergo lineage reprogramming/transdifferentiation, effectively replacing lost podocytes and restoring nephron function.• Establish primary and conditionally immortalized cell lines from renal tissue and bone.• Isolate cell populations through differential sieving, selective media and cloning rings.• Perform lineage reprogramming on primary and conditionally immortalized cell lines.• Utilize FACS to isolate transgenic cells from experimental animals.• Amplify RNA from FACS populations for qRT-PCR, NGS library preparation and microarray.• Conduct multiplexed immunofluorescence on cell monolayers and mounted tissue sections.• Create Java scripts for pre-processing of confocal images with ImageJ.• Write R scripts to conduct GO, GSEA and SPIA/GAGE analysis of microarray and NGS data.• Predict gene targets using custom data integration of multiple public datasets using R. • Validate gene targets via qRTPCR and western blot.• Perform visual data dimensionality reduction using CIRCOS (Perl) and SOM (R).• Assist in mouse uninephrectomy procedures.• Perform follow-up care on uninephrectomized mice.• Manage aging experimental murine cohorts to ensure correct study size.• Euthanize murine subjects via intraperitoneal injection or cervical dislocation according to IACUC standards.• Wrote, edited and contributed to peer-reviewed journal articles.• Contributed to slide decks for national and international scientific meetings.

Melbourne, Victoria, Australia

Performed data entry operations on behalf of the Aspirin in Reducing Events in the Elderly (ASPREE) trial. • Confirm study questionnaire completion.• Validate accuracy of machine-entered documents against hard copies.• Correct and update machine-entered database record discrepancies.• Maintain study subject confidentiality.• File study questionnaires accordingly.

Melbourne, Victoria, Australia

Aid in the study of novel anti-inflammatory lipoxins and their role in reducing diabetes-induced kidney disease and atherosclerosis.• Isolate total RNA from human and mouse urinary exosome.• Conduct necropsy of heart and abdominal and thoracic aorta.• Clean and pin full length aorta and assess atherosclerosis via oil-red-O histochemistry.• Process, mount and section frozen and paraffin embedded necropsy tissues.• Perform and quantitate immunofluorescent and immunohistochemical staining.• Ascetically prepare and deliver novel pharmaceuticals to experimental animals via intraperitoneal injection.• Carry out post-injection monitoring of treated experimental animals.

Melbourne, Australia

Studied the effect of microRNA in the onset and advancement of diabetes-induced kidney fibrosis. Work was specifically focused on the interplay of microRNA-21 and TGFβ in proximal tubule epithelial cells and how these factors influence induction of epithelial-to-mesenchymal transition. Multiple targets were bioinformatically identified and their contributions to cellular dysfunction delineated in canonical and non-canonical TGFβ signaling. Non-canonical modes of fibrotic induction, including mitochondrial dysfunction were also investigated.• Modify gene expression through transfection of oligonucleotides and gene expression vectors.• Measure growth factors secreted in culture media via ELISA.• Assess cellular respiration and mitochondrial dynamics in response to microRNA/growth factors via western blot, 2D/3D confocal microscopy, multicolor flow cytometry and Seahorse XF.• Demonstrate correlation between localized microRNA expression levels, tissue pathology and human disease.• Integrate microRNA and mRNA NGS data with KEGG pathways to generate custom networks.• Bioinformatically identify and validate novel mitochondria associated microRNA targets.• Design 3’ UTR constructs and utilize heat competent E.coli to generate luciferase expression vectors.• Delineate the contribution multiple microRNA targets to fibrotic TGFβ signaling. • Design, sub-clone, assemble and deliver shRNA-GFP and 3’UTR-GFP AAV in vivo.• Perform ultrasound microbubble-mediated delivery of shRNA expression vectors in vivo.• Train and guide junior lab members in lab skills.• Present research at international conferences and regularly give seminars at internal meetings.• Write, edit and contribute to peer-reviewed journal publications, book chapters and seminar presentations.• Execute material transfer agreements for novel cell lines and small molecule inhibitors.• Collaborate internationally to gain access to human biopsy material and aid in development of novel pharmaceuticals.

Melbourne, Victoria, Australia

Aid in collecting tissues from experimental mice for a variety projects concerned with diabetes-induced kidney disease, heart disease, atherosclerosis and oxidative stress.• Perform necropsy of all major organs including abdominal and thoracic aorta.• Embed fresh tissues in optimal cutting temperature media in preparation for cryosectioning.• Dissect tissues accordingly and fix in prepared paraformaldehyde for paraffin embedding by external services.• Utilize phenol extraction to purify RNA from human pancreatic biopsies.• Amplify pancreatic RNA samples and assess mRNA and microRNA expression.

Melbourne, Australia

Assessed the role of microRNA-21 in diabetes-induced proximal tubule epithelial cell-mediated kidney fibrosis with a particular focus on epithelial-to-mesenchymal transition.• Designed, generated and subcloned gene expression vectors utilizing transformed E.coli and transfected mammalian cells for use in adenoviral (AAV), lentiviral and conventional transfection and transduction gene delivery systems. • Transfected microRNA mimics and nucleic acid inhibitors to demonstrate microRNA roles in growth factor induced gene expression. • Analyzed secreted growth factors in cell culture media by enzyme-linked immunosorbent assay (ELISA).• Harvested and prepared mouse tissues for further analysis including RNA analysis, protein extraction and histological assessment.• Trained incoming lab members (graduate students, MD PhDs, visiting academics) in basic laboratory skills.• Presented research at local research conferences and regularly gave seminars at internal meetings.

Melbourne, Victoria, Australia

• Generate heat competent E.coli.• Establish primary cell lines from renal tissue via density gradient centrifugation.• Aid in development of bovine serum albumin-based RNA harvesting protocol.

Brisbane, Queensland, Australia

Conducted gene promoter assays as part of a larger project concerned with the major isoforms of Receptor for Advanced Glycation End products (RAGE) and their contribution to diabetes-induced kidney disease.• Sub-clone gene promoters to luciferase expression vectors.• Bulk generate sub-cloned expression vectors utilizing heat competent E.coli.• Conduct gene promoter assays with co-transfected gene product vectors.• Consult with external stakeholders to determine project objectives.• Relay project findings and implications for parent project to external stakeholders.

Melbourne, Victoria, Australia

Assisted principal investigators in designing and sub-cloning gene expression vectors and assessing microRNA activity in a variety of kidney cell types in the setting of diabetes. • Design and sub-clone multiple 3’ UTR luciferase gene expression vectors.• Conduct luciferase assays to assess microRNA binding and translational repression of expression constructs.• Assess microRNA targets in vitro via qRTPCR and western blot of target genes.• Routinely performed RNA and protein extraction from in vitro samples.• Designed and subcloned 3’ UTR luciferase expression vectors utilizing E.coli.• Created buffers and other standard laboratory reagents according to required specifications and laboratory standards.• Gained familiarity with qPCR, western blot, FACS, mammalian cell culture, bacterial cell culture, sterile technique, gene delivery (transduction, transfection and transformation).

Melbourne, Victoria, Australia

Assist principal investigators with the design and construction of lentiviral gene expression vectors, the assembly of virus particles and the establishment of lentivirus transduced cell lines.• Aid design of a tetracyclin activated microRNA lentiviral expression vector containing an IRES controlled GFP gene.• Aid in sub-cloning of expression vector components and transform E.coli via electroporation• Conduct lentiviral packaging utilizing 293T cells.• Perform dual resistance marker clonal selection of transformed E.coli.• Conduct clonal selection of lentivirus transduced cell lines and confirm transgene activity by qRTPC and fluorescent microscopy.

Doctor Of Philosophy - Phd, Biochemistry And Molecular Biology

Activities and Societies: Student Committee Vice-President (2013)Thesis title: "microRNA-21 in Diabetic Nephropathy and a Cross-Sectional.

Bachelor Of Science Honors, High Distinction (4.0 Gpa)

Activities and Societies: Golden Key International Honour Society; Deans Honour Role (2011)

Bachelor Of Biotechnoloy And Molecular Cell Biology, Biochemistry And Molecular Biology, High Distinction (4.0 Gpa)

Activities and Societies: Golden Key International Honour Society; Dean's Honour Role (2008, 2009, 2010); LUMC (La Trobe Mountaineering.