Progress report December 2020 - May 2021
Since the previous progress update (infographic) a number of objectives have been completed, and work is underway with the other aims outlined in the PhD project.
1. 16S rRNA data has been analysed (microbial community profiling)
Faecal samples were collected from dogs with Cutaneous and Renal Glomerular Vasculopathy (CRGV) and from healthy controls and sequenced using 16S rRNA gene sequencing. The data were analysed using a bioinformatic pipeline. The sequencing data were examined to investigate alpha- and beta-diversity. Alpha- diversity explores the total number of species present within a whole group, while beta-diversity explores the total number of species and the relative abundance of these species in individual samples. Sequencing of the faecal microbiome populations revealed groups that were overrepresented in the CRGV cohort in comparison to the healthy control cohort.
2. Processing CRGV and healthy control samples via 1H Nuclear Magnetic Resonance (1H NMR) (metabolic profiling)
The metabolic profile of CRGV affected dogs is being investigated to complement the microbial profiling of the gut microbiota. The metabolic signature of each sample contains thousands of molecular components that reflect biochemical events occurring within the gut microbiota, and we hope to see whether there is a characteristic metabolic profile associated with CRGV. Hopefully, this will prove effective in identifying novel biomarkers (e.g. glucose and short-chain fatty acids) associated with disease onset and potential disease susceptibility. Analysis of faecal water samples is the first aspect of this study and is being carried out in collaboration with a partner institution.
3. Clinical pathology- determining the presence of Anti-neutrophil Cytoplasmic Antibodies (ANCAs) as a potential predictor for CRGV
Enzyme-linked immunosorbent assay (ELISA) has been used to detect ANCAs in serum samples from affected and case-control dogs. ANCAs are being investigated because post-mortem examination of tissues from CRGV patients has previously revealed vasculitis of the glomerular arteries and surrounding tissue, typical hallmarks of ANCA associated vasculitis (AAV). ANCAs is a variable serological marker associated with vasculitis in a heterogeneous group of disorders characterised by inflammation and necrosis of small- to medium-sized blood vessels. Therefore, ANCAs could be used as markers for disease prediction and/ or monitoring disease progression.
In summary, over the past 6 months the study has:
Since the previous progress update (infographic) a number of objectives have been completed, and work is underway with the other aims outlined in the PhD project.
1. 16S rRNA data has been analysed (microbial community profiling)
Faecal samples were collected from dogs with Cutaneous and Renal Glomerular Vasculopathy (CRGV) and from healthy controls and sequenced using 16S rRNA gene sequencing. The data were analysed using a bioinformatic pipeline. The sequencing data were examined to investigate alpha- and beta-diversity. Alpha- diversity explores the total number of species present within a whole group, while beta-diversity explores the total number of species and the relative abundance of these species in individual samples. Sequencing of the faecal microbiome populations revealed groups that were overrepresented in the CRGV cohort in comparison to the healthy control cohort.
2. Processing CRGV and healthy control samples via 1H Nuclear Magnetic Resonance (1H NMR) (metabolic profiling)
The metabolic profile of CRGV affected dogs is being investigated to complement the microbial profiling of the gut microbiota. The metabolic signature of each sample contains thousands of molecular components that reflect biochemical events occurring within the gut microbiota, and we hope to see whether there is a characteristic metabolic profile associated with CRGV. Hopefully, this will prove effective in identifying novel biomarkers (e.g. glucose and short-chain fatty acids) associated with disease onset and potential disease susceptibility. Analysis of faecal water samples is the first aspect of this study and is being carried out in collaboration with a partner institution.
3. Clinical pathology- determining the presence of Anti-neutrophil Cytoplasmic Antibodies (ANCAs) as a potential predictor for CRGV
Enzyme-linked immunosorbent assay (ELISA) has been used to detect ANCAs in serum samples from affected and case-control dogs. ANCAs are being investigated because post-mortem examination of tissues from CRGV patients has previously revealed vasculitis of the glomerular arteries and surrounding tissue, typical hallmarks of ANCA associated vasculitis (AAV). ANCAs is a variable serological marker associated with vasculitis in a heterogeneous group of disorders characterised by inflammation and necrosis of small- to medium-sized blood vessels. Therefore, ANCAs could be used as markers for disease prediction and/ or monitoring disease progression.
In summary, over the past 6 months the study has:
- Defined a bacterial community profile associated with CRGV affected dogs.
- 1H NMR has been used to study the metabolic profile of CRGV samples.
- ANCAs have been investigated in CRGV and healthy serum samples.
Update from from Jack Whitehouse, PhD researcher, January 2021
Jack Whitehouse, PhD researcher - introduction post
Jack, the new PhD researcher has sent the attached introduction to himself now that he has started in his new role.
Jack, the new PhD researcher has sent the attached introduction to himself now that he has started in his new role.
| jack_whitehouse_post_07.05.2020.pdf |
Three year PhD studentship - The role of the microbiome and circulating endothelial cells in the pathobiology of cutaneous renal glomerular vasculopathy (CRGV)
For further detail on this funding please click on the link below
3_year_phd_studentship_crgv_project_2020_-_arrf_website_summary__pdf_.pdf
3_year_phd_studentship_crgv_project_2020_-_arrf_website_summary__pdf_.pdf