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MICB 475: Data Science Research in Microbiology - Team 2 - Lab Notebook

Team Members - Ali, Alicia, Cayden, David, John

Link to dataset source from Cirstea et al. - https://doi.org/10.1002/mds.28052

Project Summary

Parkinson’s Disease (PD) is a neurodegenerative disorder associated with dopaminergic neuron loss, leading to dopamine dysregulation. Dopaminergic therapeutics are often administered to restore dopamine levels and have been associated with changes to the gut microbiota. Through a secondary data analysis of a cross-sectional cohort of PD patients, we aimed to explore the use of four dopaminergic drugs (entacapone, pramipexole, rasagiline, amantadine) and the associated changes in the gut microbiome. Although the use of dopaminergic therapeutics was not associated with compositional alterations to the microbial diversity of PD patients, we observed changes to specific taxa. Amantadine and pramipexole therapeutics were both associated with a core microbiome that contains Faecalibacterium – a genus contained in the core microbiome of healthy individuals but absent in untreated PD patients. Furthermore, entacapone and amantadine use was associated with taxa that are indicative of a healthy gut microbiome, including Lachnospiraceae and Colidextribacter. We also identified three genera that were differentially abundant with dopaminergic drug use. Dopaminergic therapeutic use was generally associated with increased Bifidobacterium, decreased Prevotella, and increased Akkermansia. While increased Bifidobacterium is associated with a healthier gut microbiome and Akkermansia is associated with gut dysbiosis, the effects of Prevotella remain unclear. Our findings suggest that dopaminergic therapeutics are associated with alterations in the gut microbiome of PD patients that provide an overall benefit to the host. Future studies could incorporate higher resolution analysis at the species level, and explore causational effects of dopaminergic drugs in a prospective study.

Project Aims with Links to Annotated Scripts

Click on the aim to view output files
Aim 1A - QIIME2 processing
Aim 1B - Metadata wrangling

Aim 2 - Alpha and beta diversity analysis in R

Aim 3 - Core microbiome analysis

Aim 4 - Indicator species analysis

Aim 5 - Differential abundance analysis

Project Timeline

image

Meeting Notes

Final Consolidated Meeting Notes
Individual Meeting Notes - All

February

March

April

Lab notebook

Includes brief summaries of what was completed throughout the project duration.

April 15, 2024

Alicia - updated core microbiome

April 11, 2024

Cayden - updated volcano plot titles

April 6, 2024

Ali - updated the diversity plots

April 3, 2024

Alicia - updated the color codes for core microbiome figures

March 31, 2024

Cayden - updated DESeq plots

March 30, 2024

Ali - Linear regression analysis of identified genera from DESeq

March 25, 2024

David - Added meeting notes of all previous meetings up to this point Ali - Added additional files Alicia - Added abundances for core microbiome along with a 3 way venn diagram

March 24, 2024

David - Minor changes to ISA

March 23, 2024

Cayden - minor changes to plots

March 22, 2024

Cayden - Updated the genus bar plots

March 21, 2024

David - Changed ISA output, adjusted and fixed some code Cayden - Added genus sums for DESeq analysis

March 20, 2024

Cayden - updated DESeq barplot titles

March 18, 2024

Cayden - Reformatted barplots Alicia - reconducted core microbiome analysis at 1% detection

March 17, 2024

Cayden:

  • uploaded optional code for volcano plots including labels
  • further DESeq analysis results
  • including excel sheets (csvs) for comparison across all the genuses

John - changed formatting

March 16, 2024

Cayden - conducted DESeq analysis and uploaded code for DESeq, generated figures and plots John - fixing project files and file paths for DESeq

March 12, 2024

Ali - conducted pairwise beta diversity analysis and comparisons

March 11, 2024

David - Changed code for ISA, added excel sheets to generate tables for manuscript

March 10, 2024

Alicia - adding to core microbiome code and providing venn diagram images as potential figures

March 9, 2024

Ali - addded the alpha and beta diversity metrics, along with code

March 7, 2024

David - Updated script for indicator species analysis, added R script for ISA, edited some code

March 4, 2024

Ali - Added Meeting agenda, core microbiome analysis

March 3, 2024

Alicia - Added TSV and code for rough heatmap

February 12, 2024

David - Edited the R file, moved into R folder

John - fixed R file path script

Ali - Changed 'treatment' column from numeric to factor; saved metadata as .txt file; created new RProject for phyloseq object; created script for phyloseq object 'pd_phyloseq' and saved as .RData file

John - Uploaded new pd_metadata_treatment.tsv to server. Compiled table, taxa bar plots, and no mitochondria no chloroplast table as viewable .qzv files with new pd_metadata_treatment.tsv file.

John - Using table-no-mitochondria-no-chloroplast_treatment.qzv, we found a maximum sampling depth of 5421 in order to retain at least 6 samples in all treatment categories. Alpha-rarefaction curve displayed that 5421 falls under the plateau.

February 11, 2024

John - Created README file. Imported and demultiplexed parkinsons sequences. Denoising, clusterng, taxonomic training, filtering, and export conducted.

Ali - Removed NAs from PD drug columns. Added "treatment" column numbered 1-7 for the following treatments:
control = 1
pd_untreated = 2
pd_entac = 3
pd_prami = 4
pd_rasag = 5
pd_amant = 6
pd_combo = 7

Visualization of sequences on view.qiime.org

John - All high quality reads throughout. Setting a standard of 30 for median quality score, no trimming is required. 251 is the trimming parameter and the length of all of the sequences.

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