Diane K. Adams

Circulating Cancer-Associated Macrophage-Like Cells Differentiate Malignant Breast Cancer and Benign Breast Conditions

Background: Blood-based testing can be used as a noninvasive method to recover and analyze circulating tumor-derived cells for clinical use. Circulating cancer-associated macrophage-like cells (CAML) are specialized myeloid cells found in peripheral blood and associated with the presence of solid malignancies. We measured CAMLs prospectively in peripheral blood to ascertain their prevalence, specificity, and sensitivity in relation to breast disease status at clinical presentation. Methods: We report on two related but separate studies: 1) CellSieve microfilters were used to isolate CAMLs from blood samples of patients with known malignant disease (n = 41). Prevalence and specificity was compared against healthy volunteers (n = 16). 2) A follow-up double-blind pilot study was conducted on women (n = 41) undergoing core-needle biopsy to diagnose suspicious breast masses. Results: CAMLs were found in 93% of known malignant patients (n = 38/41), averaging 19.4 cells per sample, but none in the healthy controls. In subjects undergoing core biopsy for initial diagnosis, CAMLs were found in 88% of subjects with invasive carcinoma (n = 15/17) and 26% with benign breast conditions (n = 5/19). Conclusion: These preliminary pilot studies suggest that the presence of CAMLs may differentiate patients with malignant disease, benign breast conditions, and healthy individuals. Impact: We supply evidence that this previously unidentified circulating stromal cell may have utility as a screening tool to detect breast cancer in various malignancies, irrespective of disease stage. Cancer Epidemiol Biomarkers Prev; 25(7); 1037–42. ©2016 AACR.

Sequential tracking of PD-L1 expression and RAD50 induction in circulating tumor and stromal cells of lung cancer patients undergoing radiotherapy

Purpose: Evidence suggests that PD-L1 can be induced with radiotherapy and may be an immune escape mechanism in cancer. Monitoring this response is limited, as repetitive biopsies during therapy are impractical, dangerous, and miss tumor stromal cells. Monitoring PD-L1 expression in both circulating tumor cells (CTCs) and circulating stromal cells (CStCs) in blood-based biopsies might be a practical alternative for sequential, noninvasive assessment of changes in tumor and stromal cells. Experimental Design: Peripheral blood was collected before and after radiotherapy from 41 patients with lung cancer, as were primary biopsies. We evaluated the expression of PD-L1 and formation of RAD50 foci in CTCs and a CStC subtype, cancer-associated macrophage-like cells (CAMLs), in response to DNA damage caused by radiotherapy at the tumor site. Results: Only 24% of primary biopsies had sufficient tissue for PD-L1 testing, tested with IHC clones 22c3 and 28-8. A CTC or CAML was detectable in 93% and 100% of samples, prior to and after radiotherapy, respectively. RAD50 foci significantly increased in CTCs (>7×, P < 0.001) and CAMLs (>10×, P = 0.001) after radiotherapy, confirming their origin from the radiated site. PD-L1 expression increased overall, 1.6× in CTCs (P = 0.021) and 1.8× in CAMLs (P = 0.004): however, individual patient PD-L1 expression varied, consistently low/negative (51%), consistently high (17%), or induced (31%). Conclusions: These data suggest that RAD50 foci formation in CTCs and CAMLs may be used to track cells subjected to radiation occurring at primary tumors, and following PD-L1 expression in circulating cells may be used as a surrogate for tracking adaptive changes in immunotherapeutic targets. Clin Cancer Res; 23(19); 5948–58. ©2017 AACR.

Mitosis in circulating tumor cells stratifies highly aggressive breast carcinomas.

BackgroundEnumeration of circulating tumor cells (CTCs) isolated from the peripheral blood of breast cancer patients holds promise as a clinically relevant, minimally invasive diagnostic test. However, CTC utility has been limited as a prognostic indicator of survival by the inability to stratify patients beyond general enumeration. In comparison, histological biopsy examinations remain the standard method for confirming malignancy and grading malignant cells, allowing for cancer identification and then assessing patient cohorts for prognostic and predictive value. Typically, CTC identification relies on immunofluorescent staining assessed as absent/present, which is somewhat subjective and limited in its ability to characterize these cells. In contrast, the physical features used in histological cytology comprise the gold standard method used to identify and preliminarily characterize the cancer cells. Here, we superimpose the methods, cytologically subtyping CTCs labeled with immunohistochemical fluorescence stains to improve their prognostic value in relation to survival.MethodsIn this single-blind prospective pilot study, we tracked 36 patients with late-stage breast cancer over 24 months to compare overall survival between simple CTC enumeration and subtyping mitotic CTCs. A power analysis (1-β = 0. 9, α = 0.05) determined that a pilot size of 30 patients was sufficient to stratify this patient cohort; 36 in total were enrolled.ResultsOur results confirmed that CTC number is a prognostic indicator of patient survival, with a hazard ratio 5.2, p = 0.005 (95 % CI 1.6–16.5). However, by simply subtyping the same population based on CTCs in cytological mitosis, the hazard ratio increased dramatically to 11.1, p < 0.001 (95 % CI 3.1–39.7).ConclusionsOur data suggest that (1) mitotic CTCs are relativity common in aggressive late-stage breast cancer, (2) mitotic CTCs may significantly correlate with shortened overall survival, and (3) larger and more defined patient cohort studies are clearly called for based on this initial pilot study.

Synergistic negative effects of thermal stress and altered food resources on echinoid larvae

Multiple changes to the marine environment under climate change can have additive or interactive (antagonistic or synergistic) effects on marine organisms. Prompted by observations of anomalously warm sea temperatures and low chlorophyll concentrations during the 2013–2016 warm “Blob” event in the Northeast Pacific Ocean, we examined the combined effects of thermal stress and a shift in food resources on the development of a larval echinoid (Strongylocentrotus droebachiensis) in the laboratory. A high concentration of phytoplankton yielded faster echinus rudiment development at warm versus historical temperature, indicating a mitigating effect of abundant food on thermal stress; however, low phytoplankton concentration or a shift in diet to suspended kelp detritus, yielded slow development and high mortality at warm temperature. The results indicate a synergistic negative effect of thermal stress and altered food resources on larvae of a keystone marine species.

Effect of coastal marine protection on childhood health: an exploratory study

Abstract Background The integrity of ocean ecosystems are currently under threat from a suite of anthropogenic drivers including climate change, over-fishing, land-based pollution, and resource exploitation. Recent research has shown that this degradation is likely to lead to negative, long-term livelihood, biodiversity, and economic impacts. In view of the level of dependence of those in the developing countries on well-functioning ecosystems, already marginalised coastal populations are likely to suffer most of the costs of the degradation of coastal and marine ecosystems. One policy intervention that has been touted to deliver on conservation and longer-term development goals is the continued establishment of marine protected areas (MPAs). While preliminary research has shown mixed results on the effects of MPAs, some studies show, in certain contexts, positive benefits of MPAs flowing to impoverished local populations. Methods Here, we used a three-step process to see if we could detect an association between creation of MPAs and particular human health outcomes. We used childhood stunting as the dependent measure of human health. We built a database of 47 992 children living less than 25 km from a marine coast in 25 developing countries. We combined socioeconomic and health data from Demographic and Health Surveys with available climate and environmental data to examine this relationship. For analysis, we first used an information-theoretic approach to examine the potential association between distance to MPA and childhood stunting, while controlling for a suite of covariates and potential confounding socioeconomic variables. Second, we used a mixed-effect logit model to test proximity to MPAs and severe stunting in children. Third, we used propensity score matching to test the treatment effect of an MPA further while controlling for the same environmental and socioeconomic factors as in the logit models. Findings We find that the distance to MPA does show up in the top models using an information-theoretic approach. With the logit models, we find that the further from an MPA a child lives, the greater the chance a child has of being severely stunted (p Interpretation While much work needs to be done to uncover a potential causal link between well-functioning marine ecosystems and childhood health, our results indicate that such an examination might prove fruitful, and ultimately that marine conservation could be a key mechanism to improve the health of the millions of marginalised coastal peoples worldwide. Funding The Oceans and Fisheries Initiative at the Rockefeller Foundation and the National Socio-Environmental Synthesis Center.

Divergent evolutionary histories of DNA markers in a Hawaiian population of the coral Montipora capitata

We investigated intra- and inter-colony sequence variation in a population of the dominant Hawaiian coral Montipora capitata by analyzing marker gene and genomic data. Ribosomal ITS1 regions showed evidence of a reticulate history among the colonies, suggesting incomplete rDNA repeat homogenization. Analysis of the mitochondrial genome identified a major (M. capitata) and a minor (M. flabellata) haplotype in single polyp-derived sperm bundle DNA with some colonies containing 2–3 different mtDNA haplotypes. In contrast, Pax-C and newly identified single-copy nuclear genes showed either no sequence differences or minor variations in SNP frequencies segregating among the colonies. Our data suggest past mitochondrial introgression in M. capitata, whereas nuclear single-copy loci show limited variation, highlighting the divergent evolutionary histories of these coral DNA markers.