Guidelines for Transparency on Gut Microbiome Studies in Essential and Experimental Hypertension

Abstract

Hypertension is a complex and modifiable condition in which environmental factors contribute to both onset and progression. Recent evidence has accumulated for roles of diet and the gut microbiome as environmental factors in blood pressure regulation. However, this is complex because gut microbiomes are a unique feature of each individual reflecting that individual’s developmental and environmental history creating caveats for both experimental models and human studies. Here, we describe guidelines for conducting gut microbiome studies in experimental and clinical hypertension. We provide a complete guide for authors on proper design, analyses, and reporting of gut microbiota/microbiome and metabolite studies and checklists that can be used by reviewers and editors to support robust reporting and interpretation. We discuss factors that modulate the gut microbiota in animal (eg, cohort, controls, diet, developmental age, housing, sex, and models used) and human studies (eg, blood pressure measurement and medication, body mass index, demographic characteristics including age, cultural identification, living structure, sex and socioeconomic environment, and exclusion criteria). We also provide best practice advice on sampling, storage of fecal/cecal samples, DNA extraction, sequencing methods (including metagenomics and 16S rRNA), and computational analyses. Finally, we discuss the measurement of short-chain fatty acids, metabolites produced by the gut microbiota, and interpretation of data. These guidelines should support better transparency, reproducibility, and translation of findings in the field of gut microbiota/microbiome in hypertension and cardiovascular disease.

A guide to human microbiome research: study design, sample collection, and bioinformatics analysis

“…[ 45 ] On the other hand, individuals who underwent medications in the preceding several months should be excluded from a microbiome study. [ 41 , 49 ] For example, a patient treated with antimicrobial drugs 3 to 6 months before a microbiome study should be excluded. [ 49 ]…”

“…[ 41 , 49 ] For example, a patient treated with antimicrobial drugs 3 to 6 months before a microbiome study should be excluded. [ 49 ]…”

Maternal exposures and the infant gut microbiome: a systematic review with meta-analysis

“…Current variability in the literature pertaining to sample size, methodologies used to characterize the microbiome, and reporting standards have also contributed to inconsistent and often non-comparable results. Best-practice guidelines for microbiome analysis and reporting provide a useful starting point from which to ground future research, 190,192 alongside tailoring research designs, such that they are powered to reliably answer these knowledge gaps. Future research should also consider exposures during the preconception environment, which are arguably equally important in establishing longterm health and disease risk.…”

The promise and challenge of cancer microbiome research

“…To deal with these challenges, several groups have provided guidelines that researchers can utilize to improve the utility and robustness of their findings, including MIMARKS and MIMAG for reporting sequencing data from microbes [208,209], STROBE and STROME-ID for molecular data tied to epidemiological projects [210,211], and REMARK for data used as tumor markers [212,213]. Marques and colleagues, working in the field of hypertension, have published a set of general guidelines and checklists that researchers and publishers can use when designing, executing, and reporting microbiome-related studies of disease in either animal models or humans that could easily be adapted to cancer research [214]. Other groups, including the International Human Microbiome Standards Project and the Microbiome Quality Control project, have proposed research and reporting guidelines for experimental design when assessing the microbiome in humans and model systems [14,215,216].…”