Elucidating the roles of the mammary and gut microbiomes in breast cancer development

The mammary microbiome is a newly characterized bacterial niche that might offer biological insight into the development of breast cancer. Together with in-depth analysis of the gut microbiome in breast cancer, current evidence using next-generation sequencing and metabolic profiling suggests compositional and functional shifts in microbial consortia are associated with breast cancer. In this review, we discuss the fundamental studies that have progressed this important area of research, focusing on the roles of both the mammary tissue microbiome and the gut microbiome. From the literature, we identified the following major conclusions, (I) There are unique breast and gut microbial signatures (both compositional and functional) that are associated with breast cancer, (II) breast and gut microbiome compositional and breast functional dysbiosis represent potential early events of breast tumor development, (III) specific breast and gut microbes confer host immune responses that can combat breast tumor development and progression, and (IV) chemotherapies alter the microbiome and thus maintenance of a eubiotic microbiome may be key in breast cancer treatment. As the field expectantly advances, it is necessary for the role of the microbiome to continue to be elucidated using multi-omic approaches and translational animal models in order to improve predictive, preventive, and therapeutic strategies for breast cancer

Acute Beetroot Juice Ingestion Increases Nitric Oxide Bioavailability Without Changing Oral Microbial Composition in Healthy Young Women

Dietary nitrate supplementation can elicit beneficial health and exercise performance effects. Oral microbiota are critical for the metabolism of exogenously consumed nitrate; however, limited data are available on the influence of dietary nitrate ingestion on bacterial taxa and in women. PURPOSE: To investigate if acute dietary nitrate ingestion alters the oral microbiota in young healthy women compared to a nitrate-depleted placebo. METHODS: In a randomized double-blinded crossover design, fifteen recreationally active women (mean ± SD: age 20 ± 1 years; body mass 63 ± 10 kg; height 1.68 ± 0.1 m) participated in two conditions to ingest nitrate-rich beetroot juice (BR; 12 mmol of nitrate) and nitrate-depleted beetroot juice (PL, negligible nitrate), 2.5 hours prior to a resting blood draw and buccal swab sample. Plasma [nitrate] and [nitrite] were analyzed using gas phase chemiluminescence. Buccal swab samples were used for DNA extraction and isolation. DNA was amplified using polymerase chain reaction targeting the V3 - V4 region of the 16S rRNA gene. Following index PCR, amplicons were pooled and sequenced using the iSeq Illumina NGS sequencer. Reads were clustered into amplicon sequence variants and analyzed for alpha and beta diversity and relative abundance. RESULTS: BR increased plasma [nitrate] (PL: 52 ± 14 µM vs. BR: 629 ± 132 µM, P \u3c 0.001) and plasma [nitrite] (PL: 276 ± 286 nM vs. BR: 703 ± 391 nM, P \u3c 0.001). One sample had insufficient DNA and thus, a subset of samples was analyzed for oral microbial composition (n = 14). Alpha (i.e., species richness or evenness) and beta diversity was not different between PL and BR (P \u3e 0.05). The relative abundance of the phylum and genus were not influenced by BR (P \u3e 0.05). CONCLUSION: Acute nitrate ingestion did not improve or worsen the composition of global or lower taxonomic levels of bacteria in young recreationally active women. These data indicate that acute nitrate ingestion is an intervention to rapidly increase nitric oxide bioavailability in young recreationally active women, which is an effect that did not require changes to the oral microbial community. Further research is required to understand the impact of dosing regimen and population on oral bacterial taxa and the efficacy of nitrate on nitrate-induced effects

Composition and Functional Potential of the Human Mammary Microbiota Prior to and following Breast Tumor Diagnosis

Microbiota studies have reported changes in the microbial composition of the breast upon cancer development. However, results are inconsistent and limited to the later phases of cancer development (after diagnosis). We analyzed and compared the resident bacterial taxa of histologically normal breast tissue (healthy, H, n = 49) with those of tissues donated prior to (prediagnostic, PD, n = 15) and after (adjacent normal, AN, n = 49, and tumor, T, n = 46) breast cancer diagnosis (n total = 159). DNA was isolated from tissue samples and submitted for Illumina MiSeq paired-end sequencing of the V3-V4 region of the 16S gene. To infer bacterial function in breast cancer, we predicted the functional bacteriome from the 16S sequencing data using PICRUSt2. Bacterial compositional analysis revealed an intermediary taxonomic signature in the PD tissue relative to that of the H tissue, represented by shifts in Bacillaceae, Burkholderiaceae, Corynebacteriaceae, Streptococcaceae, and Staphylococcaceae. This compositional signature was enhanced in the AN and T tissues. We also identified significant metabolic reprogramming of the microbiota of the PD, AN, and T tissue compared with the H tissue. Further, preliminary correlation analysis between host transcriptome profiling and microbial taxa and genes in H and PD tissues identified altered associations between the human host and mammary microbiota in PD tissue compared with H tissue. These findings suggest that compositional shifts in bacterial abundance and metabolic reprogramming of the breast tissue microbiota are early events in breast cancer development that are potentially linked with cancer susceptibility

Co-Ingestion of Dietary Nitrate and Ascorbic Acid on Nitric Oxide Biomarkers and The Oral Microbiome in Sedentary Hispanic Women

Nitric oxide bioavailability increases following nitrate supplementation wherein oral microbiota facilitate the metabolism and absorption of nitrate. However, few studies have examined if co-ingestion of nitrate with antioxidants can further elevate nitric oxide bioavailability. Moreover, our understanding on how the oral microbiome responds to nitrate supplementation is limited, especially in women. PURPOSE: To examine the effects of ingesting dietary nitrate and ascorbic acid independently and concurrently on markers of nitric oxide bioavailability and oral microbiota species. METHODS: Twelve sedentary women of Hispanic descent (mean ± SD: age 20 ± 1 years; body mass 74 ± 15 kg; height 1.62 ± 0.09 m) consumed nitrate-rich beetroot juice (BR), nitrate-depleted beetroot juice (PL), ascorbic acid (AA), and crystal light (CRY) in four conditions: BR combined with AA (BR+AA); BR only (BR+CRY); AA only (PL+AA); and placebo-control (PL+CRY). Supplements were ingested 2.5 hours prior to a resting blood draw and buccal swab sample. Plasma [nitrate] and [nitrite] were analyzed using gas phase chemiluminescence. Buccal swab samples were used for DNA extraction and isolation. DNA was amplified using polymerase chain reaction (PCR) targeting the V3 - V4 region of the 16S rRNA gene. Following index PCR, amplicons were pooled and sequenced using the iSeq Illumina NGS sequencer. Reads were clustered into amplicon sequence variants and analyzed for alpha and beta diversity and relative abundance. RESULTS: BR increased plasma [nitrate] (BR+AA: 641 ± 252 vs. BR+CRY: 528 ± 307 vs. PL+AA: 35 ± 10 vs. PL+CRY: 35 ± 12 µM, P \u3c 0.001) and plasma [nitrite] (BR+AA: 710 ± 336 vs. BR+CRY: 578 ± 428 vs. PL+AA: 209 ± 88 vs. PL+CRY: 198 ± 82 nM, P \u3c 0.001) with no differences within BR and PL conditions. Alpha and beta diversity, and the relative abundance of higher and lower taxonomic levels were not significantly different between all conditions (P \u3e 0.05) CONCLUSION: Concurrent nitrate and AA supplementation did not elicit additional increases to nitric oxide compared to nitrate ingestion alone. Acute beetroot juice and ascorbic acid were ineffective at modulating oral microbial composition. Further research is required to understand the impact of supplementation regimen and population on the physiological effects of dietary nitrate

Asthma and the microbiome: defining the critical window in early life

Asthma is a chronic inflammatory immune disorder of the airways affecting one in ten children in westernized countries. The geographical disparity combined with a generational rise in prevalence, emphasizes that changing environmental exposures play a significant role in the etiology of this disease. The microflora hypothesis suggests that early life exposures are disrupting the composition of the microbiota and consequently, promoting immune dysregulation in the form of hypersensitivity disorders. Animal model research supports a role of the microbiota in asthma and atopic disease development. Further, these model systems have identified an early life critical window, during which gut microbial dysbiosis is most influential in promoting hypersensitivity disorders. Until recently this critical window had not been characterized in humans, but now studies suggest that the ideal time to use microbes as preventative treatments or diagnostics for asthma in humans is within the first 100 days of life. This review outlines the major mouse-model and human studies leading to characterization of the early life critical window, emphasizing studies analyzing the intestinal and airway microbiotas in asthma and atopic disease. This research has promising future implications regarding childhood immune health, as ultimately it may be possible to therapeutically administer specific microbes in early life to prevent the development of asthma in children.Medicine, Faculty ofScience, Faculty ofOther UBCMicrobiology and Immunology, Department ofPediatrics, Department ofReviewedFacult

The Role of the Microbiome in the Developmental Origins of Health and Disease

Although the prominent role of the microbiome in human health has been established, the early-life microbiome is now being recognized as a major influence on long-term human health and development. Variations in the composition and functional potential of the early-life microbiome are the result of lifestyle factors, such as mode of birth, breastfeeding, diet, and antibiotic usage. In addition, variations in the composition of the early-life microbiome have been associated with specific disease outcomes, such as asthma, obesity, and neurodevelopmental disorders. This points toward this bacterial consortium as a mediator between early lifestyle factors and health and disease. In addition, variations in the microbial intrauterine environment may predispose neonates to specific health outcomes later in life. A role of the microbiome in the Developmental Origins of Health and Disease is supported in this collective research. Highlighting the early-life critical window of susceptibility associated with microbiome development, we discuss infant microbial colonization, beginning with the maternal-to-fetal exchange of microbes in utero and up through the influence of breastfeeding in the first year of life. In addition, we review the available disease-specific evidence pointing toward the microbiome as a mechanistic mediator in the Developmental Origins of Health and Disease

Analysis of Microbial Water Contamination, Soil Microbial Community Structure, and Soil Respiration in a Collaborative First-Year Students as Scholars Program (SAS)

The persistence of college students in STEM majors after their first-year of college is approximately 50%, with underrepresented populations displaying even higher rates of departure. For many undergraduates, their first-year in college is defined by large class sizes, poor access to research faculty, and minimal standing in communities of scholars. Pepperdine University and Whittier College, funded by a National Science Foundation award to Improve Undergraduate Stem Education (NSF IUSE), partnered in the development of first-year classes specifically geared to improve student persistence in STEM and academic success. This Students as Scholars Program (SAS) engaged first-year undergraduates in scholarly efforts during their first semester in college with a careful approach to original research design and mentoring by both faculty and upperclassmen experienced in research. Courses began by introducing hypothesis formulation and experimental design partnered with the scientific focus of each course (ecological, biochemical, microbiological). Students split into research teams, explored the primary literature, designed research projects, and executed experiments over a 6–7 week period, collecting, analyzing, and interpreting data. Microbiology-specific projects included partnerships with local park managers to assess water quality and microbial coliform contamination at specified locations in a coastal watershed. In addition, students explored the impact of soil salinity on microbial community structure. Analysis of these samples included next-generation sequencing and microbiome compositional analysis via collaboration with students from an upper division microbiology course. This cross-course collaboration facilitated additional student mentoring opportunities between upperclassmen and first-year students. This approach provided first-year students an introduction to the analysis of complex data sets using bioinformatics and statistically reliable gas-exchange replicates. Assessment of the impact of this program revealed students to view the research as challenging, but confidence building as they take their first steps as biology majors. In addition, the direct mentorship of first-year students by upperclassmen and faculty was viewed positively by students. Ongoing assessments have revealed SAS participants to display a 15% increased persistence rate in STEM fields when compared to non-SAS biology majors

The effects of nitrate supplementation on oral microbiota composition in females

Dietary nitrate (NO3-) supplementation, improves cardiovascular, cognitive, and contractile function by augmenting nitric oxide (NO) biosynthesis; however, the specific taxa responsible, and whether NO3- ingestion can promote a favorable microbial composition to facilitate NO synthesis has yet to be established. To determine whether NO3- supplementation is associated with specific microbial taxa in females, buccal cell samples were collected from participants across two experiments: 1) 14 healthy adult females consumed an acute dose of NO3--depleted beetroot juice (PL) or NO3--rich beetroot juice (BR); and 2) 13 sedentary Hispanic females consumed an acute dose of PL or BR combined with ascorbic acid (VITC) or a crystal light beverage (CRY). Buccal samples were isolated for their 16s rRNA gene. DNA concentration was assessed with the Nanodrop guidelines of \u3e 10 ng/uL and A260/A280 values of 1.6 to 2.4. DNA concentration in experiment 1 (PL: 37.66±18.92; BR: 37.52±11.54 ng/uL) and experiment 2 (BR+VITC: 37.66±18.92; BR+CRY: 37.52±11.54 ng/uL; PL+VITC: 37.52±11.54; PL+CRY: 37.52±11.54) were adequate. DNA purity in experiment 1 (PL: 2.03±0.28; BR: 2.06±0.21) and experiment 2 (BR+VITC: 37.66±18.92; BR+CRY: 37.52±11.54 ng/uL; PL+VITC: 37.52±11.54; PL+CRY: 37.52±11.54) were adequate. Samples were amplified and checked using polymerase chain reaction and southern blots, respectively. Pooled samples are undergoing library sequencing to determine microbial taxa composition following NO3- supplementation