California Breast Cancer Research Program

BACKGROUND: We conducted a randomized, double-blinded pilot trial to compare the impact of two electroacupuncture (EA) regimens on co-occurring neuropsychiatric symptoms among breast cancer survivors (BCS). METHODS: BCS who self-reported cognitive impairment, fatigue, insomnia, or psychological distress were randomized (1:1) to receive ten weekly EA to target either neuropsychiatric-specific (nEA) or non-neuropsychiatric-specific (sEA) acupoints. Primary endpoints were the within-group pre-post effect sizes (Glass's Δ) in symptom severities, adjusted for multiple comparisons (p-adjusted). Outcomes were assessed using neurocognitive tests (CANTAB®), PROs (FACT-Cog, MFSI-SF, EORTC QLQ-C30), plasma biomarkers, and neuroimaging. Responders were defined by reliable change index (for objective cognition) or MCID (for PROs). RESULTS: Thirty-five were recruited, with 30 (86%) completing all sessions. The mean (±SD) age was 58.2 (±12.2) years, and 86% reported co-occurring symptoms. Following treatment, the nEA group demonstrated significant improvements in attention (T3: Δ = 0.562, T4: Δ = 0.708, both p-adjusted < 0.05) and distress (T3: Δ = 0.764, T4: Δ = 0.711, both p-adjusted < 0.05). More responders were observed after nEA treatment for objective cognition (42.9% vs 12.5%) and distress (50% vs 37.5%). nEA-treated participants showed increased gray matter volume compared to sEA (p = 0.033), which positively correlated with better attention function (r = 0.69, p = 0.020). nEA-related improvements in memory and response speed were associated with reduced connectivity in the Default Mode Network (DMN-SFG, r=-0.93, p < 0.01) and increased connectivity in the Dorsal Attention Network (DAN-SMG, r = 0.86, p < 0.001), respectively. All adverse events were grade 2 or lower. CONCLUSIONS: EA targeting neuropsychiatric-specific acupoints suggests improvements in cognition and distress symptoms in BCS, warranting validation in larger, multicenter trials. CLINICALTRIALS.GOV: NCT05283577.

BackgroundTriple-negative type of breast cancer (TNBC) has limited therapeutic options and frequently metastasizes, leading to low survival rates. Oxidative phosphorylation (OXPHOS) is a driver of TNBC metastasis, but the signaling underlying this metabolic change is poorly understood.MethodsWe performed metabolic assays and assessed migratory and metastatic potential in cells with manipulated CDCP1/mitochondrial Src signaling.ResultsWe show that the pro-metastatic cell surface protein CUB-domain containing protein 1 (CDCP1) activates Src kinase localized in mitochondria, which potently induces OXPHOS and TNBC migration. Genetic targeting of either CDCP1 or mitochondrial Src, as well as pharmacological inhibition of Src reduce OXPHOS in vitro. We further show that mitochondrial Src increases OXPHOS by stimulating Complex I activity in the electron transport chain. Importantly, rescuing Complex I activity in cells devoid of CDCP1/mitochondrial Src signaling restores both OXPHOS and migration. We also provide evidence that NAD+ pool generated by Complex I is contributing to the observed migratory phenotype. Lastly, we determined that inhibiting mitochondrial Src reduces metastasis in TNBC cells.ConclusionsBoth CDCP1 and mitochondrial Src represent potential therapeutic targets to inhibit OXPHOS-mediated TNBC metastasis.

Hormonal growth promotants (HGPs) are a class of pharmaceutical agents commonly administered to cattle in the United States to improve growth rates of the animal, alter behavior, or to improve the desired characteristics of retail cuts of meat. There is a concern that low residual concentrations of HGPs may remain in tissue after slaughter, and consumption of tissues containing these compounds may increase the risk of adverse health outcomes, including cancer. Sensitive and selective methods are necessary to assess exposure of HGPs by populations that consume meat products from animals that may have been administered HGPs. A liquid chromatography-tandem mass spectrometry method was developed and validated to detect the low-level presence of HGPs including estradiol, testosterone, estradiol benzoate, melengestrol, melengestrol acetate, progesterone, testosterone propionate, trenbolone, trenbolone acetate, and α-zearalanol in retail cuts of meat following a liquid-liquid extraction using a high pH solution with 30-50× less mass of meat required as compared to similar approaches. Good chromatographic performance and sensitivity was achieved utilizing ammonium fluoride as a mobile phase additive without the need for derivatization. Validation parameters including accuracy, precision, recovery, matrix effects, limits of detection, limits of quantitation, linear range, and stability were determined. The limits of detection ranged from 0.1 to 1.0 ng/g, depending on the compound, with adequate accuracy and precision without the need for extensive sample preparation approaches.

These data show the differentially expressed genes (DEG) from HCC38 breast cancer cell line chronically exposed to nicotine versus vehicle control. Additional data is also provided from dynamic trajectory analysis, identifying the most dynamic genes due to chronic nicotine treatment. To produce this dataset, we first performed single cell RNA sequencing from HCC38 cells chronically treated with vehicle or nicotine, followed by scanpy analysis to yield 6 discrete cell clusters at conservative resolution. We then evaluated differential gene expression between chronic nicotine and control cells for each individual cluster or in the whole sample using PyDESeq2. For dynamic trajectory analysis, Velocyto (0.6) was used to estimate the spliced and unspliced counts for each gene between chronic nicotine-treated cells and vehicle, allowing computation of gene velocities. These data are useful for analysing the expression of individual genes or gene velocities either in the whole sample or in the different clusters identified. Since the HCC38 cell line used in these experiments is heterogeneous, including cells with features of stem-like, luminal progenitor-like and more differentiated cells, this dataset allows examination of the conserved as well as disparate gene expression effects of nicotine in different breast cancer cell types. Our dataset has a great potential for re-use given the recent surge in interest surrounding the role tobacco-use plays in breast cancer progression.

BackgroundEndogenous and exogenous hormones may be present in beef. Human consumption of hormones has been linked to adverse health effects.ObjectiveTo estimate daily intake of hormonal growth promotants (HGP) from beef consumed by the US population.MethodsWe combined self-reported beef consumption information from a nationally-representative survey with concentrations of 12 HGP measured in 397 samples of retail beef/fat purchased in California. We defined typical, high, and maximum intake scenarios assuming self-reported consumed beef contained the mean, 95th percentile, and maximum concentrations of each HGP, respectively. We estimated distributions of usual (i.e., long-term) daily intake and short-term daily intake (µg/kg/day). We calculated the hazard quotient (HQ), or ratio of estimated intake to the World Health Organization’s acceptable daily intake (ADI) for the HGP.ResultsThe highest estimated HQs were found for melengestrol acetate (MGA). For usual daily intake under the typical intake scenario, no HQ exceeded 0.02 (0.00047 µg MGA/kg/day). Under the maximum intake scenario, the highest HQ was 0.29 (0.0087 µg MGA/kg/day), corresponding to the 99th percentile of intake among young boys (ages 1–5). The highest short-term intake estimates for MGA under the maximum intake scenario were the 99th percentile of intake among young girls and boys, which equaled (HQ = 1.00) or exceeded (HQ = 1.29) the ADI for MGA, respectively.ImpactHormonal growth promotants (HGP) are used to increase beef production and have been linked to adverse reproductive effects. We estimated daily intake of MGA and several other HGP using US nationally-representative beef consumption data collected between 2015–2018 and HGP concentrations in retail beef. Estimated intake was highest for young children, but estimates were generally very low compared to current health-based intake limits. However, these limits are typically based on studies in adult animals, and further study of potential adverse effects during sensitive developmental periods, such as in early life, may be warranted to ensure recommended intake limits are health-protective.

Smoking plays an underappreciated role in breast cancer progression, increasing recurrence and mortality in patients. Here, we show that S100A8/A9 innate immune signaling is a molecular mechanism that identifies smoking-related breast cancers and underlies their enhanced malignancy. In contrast to acute exposure, chronic nicotine increased tumorigenicity and reprogrammed breast cancer cells to express innate immune response genes. This required the α7 nicotinic acetylcholine receptor, which elicited dynamic changes in cell differentiation, proliferation, and expression of secreted cytokines, such as S100A8 and S100A9, as assessed by unbiased scRNA-seq. Indeed, pharmacologic or genetic inhibition of S100A8/A9-RAGE receptor signaling blocked nicotine’s tumor-promoting effects. We also discovered Syntaphilin (SNPH) as an S100A8/A9-dependent gene enriched specifically in estrogen receptor-negative (ER-) cancers from former smokers, linking this response to patient disease. Together, our findings describe a new α7 nAChR-S100A8/A9-Syntaphilin immune signaling module that drives nicotine-induced tumor progression and distinguishes smoking-related patient disease as a distinct subset of aggressive breast cancers.

To explore the hypothesis that differential exposures to estrogen active chemicals may contribute to regional disparities in cancer incidence, a comprehensive targeted and nontargeted analysis was conducted over two seasons (2020) for drinking water samples from 120 households served by 8 public water systems (4 with historically elevated breast cancer incidence) and from 15 brands of retail water. All samples were analyzed using gas and liquid chromatography with high-resolution mass spectrometry and a bioassay for estrogen receptor agonism. Target compounds included disinfection byproducts, per- and polyfluoroalkyl substances (PFAS), trace elements, and compounds selected for their possible relation to breast cancer. Over 7500 GC and LC nontargeted molecular features passed all quality control filters in each sampling season and were prioritized for identification if they were related to measured estrogen receptor agonism or were present at higher levels in areas with high breast cancer incidence (n = 1036). Benzothiazole-2-sulfonic acid, acetyl tributyl citrate, and diphenyl sulfone were among the prioritized and confirmed nontarget compounds. Nine polycyclic aromatic hydrocarbons and two ketone derivatives displayed significant negative correlations with estrogen receptor agonism. Many prioritized compounds remained unidentified, as 84.4% of the LC features and 77.5% of the GC features could not be annotated with high confidence.

BackgroundHomelessness is a growing concern in the United States, especially among people who use drugs (PWUD). The degree of material hardship among this population may be linked to worse health outcomes. PWUD experiencing homelessness in urban areas are increasingly subjected to policies and social treatment, such as forced displacement, which may worsen material hardship. It is critical to describe hardship among PWUD and examine if it is linked to health outcomes.MethodsData were collected as part of a prospective cohort study of PWUD in Los Angeles, California and Denver, Colorado (n = 476). Analysis sample size was smaller (N = 395) after selecting for people experiencing homelessness and for whom data were complete. Five indicators assessing hardship (difficulty finding food, clothing, restrooms, places to wash/shower, and shelter) in the past three months were obtained from participants at baseline and were used in latent class analysis (LCA). We chose a base latent class model after examination of global fit statistics. We then built three auxiliary models using the three-step Bolck–Croon–Hagenaars (BCH) method to test the relationship of latent class membership to several hypothesized social and health variables in this same three month time period.ResultsFit statistics, minimum classification probabilities, and ease of interpretation indicated a three-class solution for level of material difficulty. We termed these classes “High Difficulty” (n = 82), “Mixed Difficulty” (n = 215), and “Low Difficulty” (n = 98). Average classification probabilities indicated good class separability. “High Difficulty” participants had high probabilities of usually having difficulty accessing all five resources. “Mixed Difficulty” participants indicated a range of difficulty accessing all resources, with restrooms and bathing facilities being the most difficult. “Low Difficulty” participants were defined by high probabilities of never having access difficulty. In auxiliary analyses, there were significant (p < 0.05) differences in experiences of displacement, opioid withdrawal symptoms, nonfatal overdose, and violent victimization between classes.ConclusionsThis LCA indicates that among PWUD experiencing homelessness there exist distinct differences in resource access and material hardship, and that these differences are linked with political, social, substance use, and other health outcomes. We add to the literature on the relationship between poverty and health among PWUD. Policies which increase difficulty accessing necessary material resources may negatively impact health in this population.

Water is an increasingly precious resource in California as years of drought, climate change, pollution, as well as an expanding population have all stressed the state's drinking water supplies. Currently, there are increasing concerns about whether regulated and unregulated contaminants in drinking water are linked to a variety of human-health outcomes particularly in socially disadvantaged communities with a history of health risks. To begin to address this data gap by broadly assessing contaminant mixture exposures, the current study was designed to collect tapwater samples from communities in Gold Country, the San Francisco Bay Area, two regions of the Central Valley (Merced/Fresno and Kern counties), and southeast Los Angeles for 251 organic chemicals and 32 inorganic constituents. Sampling prioritized low-income areas with suspected water quality challenges and elevated breast cancer rates. Results indicated that mixtures of regulated and unregulated contaminants were observed frequently in tapwater throughout the areas studied and the types and concentrations of detected contaminants varied by region, drinking-water source, and size of the public water system. Multiple exceedances of enforceable maximum contaminant level(s) (MCL), non-enforceable MCL goal(s) (MCLG), and other health advisories combined with frequent exceedances of benchmark-based hazard indices were also observed in samples collected in all five of the study regions. Given the current focus on improving water quality in socially disadvantaged communities, our study highlights the importance of assessing mixed-contaminant exposures in drinking water at the point of consumption to adequately address human-health concerns (e.g., breast cancer risk). Data from this pilot study provide a foundation for future studies across a greater number of communities in California to assess potential linkages between breast cancer rates and tapwater contaminants.

The Cover Feature shows plasticware for biomarker measurement, which constitutes a notable portion of the plastic waste stream from medical consumables. We have developed a portable vortex fluidic device (P‐VFD) with a membrane array to initiate fast reaction, detection and analysis of a clinically relevant protein biomarker, p75ECD. It shows significant potential to replace traditional assay plates with a membrane, providing a greener and more sustainable alternative in biofluid biomarker measurement. More information can be found in the Research Article by X. Luo, M.‐L. Rogers, C. L. Raston and co‐workers.