Perceived Health Needs of Church Leaders and Members: A Secondary Data Analysis of the Mid-South Congregational Health Survey.

Needs assessments have been successful in helping communities and congregations focus their health ministry efforts; however, most have used leader perceptions of congregational health needs. The purpose of this study was to examine and compare the self-reported needs of both church leaders and members to be addressed by their congregation. Church leaders (n = 369) and members (n = 459) from 92 congregations completed the 2019 Mid-South Congregational Health Survey. Frequencies and generalized linear mixed models (GLMM) were performed to examine the top 10 self-reported needs and associations by church role, respectively. Of the top 10 congregational needs, anxiety or depression, high blood pressure, stress, and healthy foods were ranked identically regardless of church role. Church leaders perceived obesity and diabetes to be important congregational health needs, whereas members perceived affordable health care and heart disease to be important congregational health needs. GLMM, controlling for within-church clustering and covariates, revealed church leaders were more likely than members to report obesity (odds ratio [OR]: 1.93, 95% confidence interval [CI] = [1.39, 2.67], p < .0001) and diabetes (OR: 1.73, 95% CI = [1.24, 2.41], p = .001) as congregational needs. Findings display similarities and differences in needs reported by church role. Including many perspectives when conducting congregational health needs assessments will assist the development of effective faith-based health promotion programs.

Fit with Faith: An Exploratory Study Examining a Behavior Change Intervention for African-American Clergy and Their Spouses.

Fit with Faith is a 10-week, diet, physical activity, and stress reduction intervention for African-American clergy and spouses, which included: meetings, phone calls, a behavior tracking app. Survey, 24-h recall, accelerometer, anthropometric, and blood pressure data were collected. Wilcoxon signed ranked tests were used for analyses. In this one-arm study, clergy and spouses (n = 20) attended most meetings and calls, but only half posted daily goals or tracked behaviors using the app. Spouses' body mass index (BMI) decreased and physical activity self-regulation cognitive scores increased pre-post intervention. Statistically significant changes in BMI, systolic blood pressure, and self-regulations scores also were seen among younger (< 51 years) participants (n = 8). As positive changes were seen mostly among women and younger participants, more research is needed on how to engage all clergy in behavior change programs.

Discovery of selective small-molecule HDAC6 inhibitor for overcoming proteasome inhibitor resistance in multiple myeloma.

Multiple myeloma (MM) has proven clinically susceptible to modulation of pathways of protein homeostasis. Blockade of proteasomal degradation of polyubiquitinated misfolded proteins by the proteasome inhibitor bortezomib (BTZ) achieves responses and prolongs survival in MM, but long-term treatment with BTZ leads to drug-resistant relapse in most patients. In a proof-of-concept study, we previously demonstrated that blocking aggresomal breakdown of polyubiquitinated misfolded proteins with the histone deacetylase 6 (HDAC6) inhibitor tubacin enhances BTZ-induced cytotoxicity in MM cells in vitro. However, these foundational studies were limited by the pharmacologic liabilities of tubacin as a chemical probe with only in vitro utility. Emerging from a focused library synthesis, a potent, selective, and bioavailable HDAC6 inhibitor, WT161, was created to study the mechanism of action of HDAC6 inhibition in MM alone and in combination with BTZ. WT161 in combination with BTZ triggers significant accumulation of polyubiquitinated proteins and cell stress, followed by caspase activation and apoptosis. More importantly, this combination treatment was effective in BTZ-resistant cells and in the presence of bone marrow stromal cells, which have been shown to mediate MM cell drug resistance. The activity of WT161 was confirmed in our human MM cell xenograft mouse model and established the framework for clinical trials of the combination treatment to improve patient outcomes in MM.

Selective inhibition of BET bromodomains.

Epigenetic proteins are intently pursued targets in ligand discovery. So far, successful efforts have been limited to chromatin modifying enzymes, or so-called epigenetic 'writers' and 'erasers'. Potent inhibitors of histone binding modules have not yet been described. Here we report a cell-permeable small molecule (JQ1) that binds competitively to acetyl-lysine recognition motifs, or bromodomains. High potency and specificity towards a subset of human bromodomains is explained by co-crystal structures with bromodomain and extra-terminal (BET) family member BRD4, revealing excellent shape complementarity with the acetyl-lysine binding cavity. Recurrent translocation of BRD4 is observed in a genetically-defined, incurable subtype of human squamous carcinoma. Competitive binding by JQ1 displaces the BRD4 fusion oncoprotein from chromatin, prompting squamous differentiation and specific antiproliferative effects in BRD4-dependent cell lines and patient-derived xenograft models. These data establish proof-of-concept for targeting protein-protein interactions of epigenetic 'readers', and provide a versatile chemical scaffold for the development of chemical probes more broadly throughout the bromodomain family.

The rates, perceptions, and willingness of men who have sex with men to donate blood.

BACKGROUND: Since 1983 in the United States, any man who has had sex with another man (MSM) at any time since 1977 has been deferred from donating blood for life. Although there has been a push to change the deferral, there is a paucity of information on both the rates of MSM blood donation and the willingness of MSMs to donate if the deferral were changed. STUDY DESIGN AND METHODS: A 15-question survey was given at two lesbian, gay, bisexual, and transgender festivals in Chicago and New Orleans. Participants were asked about a previous history of blood donation and whether they would be willing to donate were the lifetime deferral changed. Participants were also asked to determine whether it was safe for hypothetical MSMs with varying sexual practices to donate blood and whether they believed that it was safe for them to donate their own blood. RESULTS: Our study found that 42.0% of all participants had not complied with the deferral policy and have donated blood at least once, with a mean number of donations of 4.84. Additionally, 85.9% of participants would be willing to donate blood if the deferral were changed. CONCLUSION: Despite the lifetime deferral, many MSMs have previously donated blood, and many more are willing to donate. Given this, along with the safe implementation of temporary deferral policies in other nations, the United States should consider adopting a temporary deferral policy for MSMs.

Model-directed engineering of "difficult-to-express" monoclonal antibody production by Chinese hamster ovary cells.

Despite improvements in volumetric titer for monoclonal antibody (MAb) production processes using Chinese hamster ovary (CHO) cells, some "difficult-to-express" (DTE) MAbs inexplicably reach much lower process titers. These DTE MAbs require intensive cell line and process development activity, rendering them more costly or even unsuitable to manufacture. To rapidly and rationally identify an optimal strategy to improve production of DTE MAbs, we have developed an engineering design platform combining high-yielding transient production, empirical modeling of MAb synthesis incorporating an unfolded protein response (UPR) regulatory loop with directed expression and cell engineering approaches. Utilizing a panel of eight IgG1 λ MAbs varying >4-fold in volumetric titer, we showed that MAb-specific limitations on folding and assembly rate functioned to induce a proportionate UPR in host CHO cells with a corresponding reduction in cell growth rate. Derived from comparative empirical modeling of cellular constraints on the production of each MAb we employed two strategies to increase production of DTE MAbs designed to avoid UPR induction through an improvement in the rate/cellular capacity for MAb folding and assembly reactions. Firstly, we altered the transfected LC:HC gene ratio and secondly, we co-expressed a variety of molecular chaperones, foldases or UPR transactivators (BiP, CypB, PDI, and active forms of ATF6 and XBP1) with recombinant MAbs. DTE MAb production was significantly improved by both strategies, although the mode of action was dependent upon the approach employed. Increased LC:HC ratio or CypB co-expression improved cell growth with no effect on qP. In contrast, BiP, ATF6c and XBP1s co-expression increased qP and reduced cell growth. This study demonstrates that expression-engineering strategies to improve production of DTE proteins in mammalian cells should be product specific, and based on rapid predictive tools to assess the relative impact of different engineering interventions.

A Scoping Review of Self-Care Within the Context of Obesity-Related Outcomes Among Faith Leaders.

OBJECTIVE: Faith leaders often serve as health-related role models yet many struggle with obesity and self-care engagement. The purpose of this scoping review was to examine how the faith leader literature has defined self-care and examined obesity and obesity-related chronic disease. DATA SOURCE: Studies were identified through database (eg, PubMed, CINAHL, PsycINFO), backward, and grey literature (eg, dissertations) searches. INCLUSION/EXCLUSION CRITERIA: Studies published in English with participants who were 18 years or older and examined leaders across all faiths. Studies also included an examination of self-care behaviors among faith leaders within the context of obesity or obesity-related chronic diseases. DATA EXTRACTION/SYNTHESIS: Data synthesis was qualitative and informed by the six-step framework developed by Arksey and O'Malley (2005) as well as updated recommendations by Daudt et al (2013). Of the 418 studies identified and screened, 20 met the eligibility criteria. RESULTS: Studies were primarily cross-sectional and participants Christian faith-leaders in the US. Most studies did not define self-care or incorporate theory, but focused on vegetarian diets and physical activity engagement. Other self-care related behaviors (eg, sleep, days off), some unique to faith leaders (eg, sabbatical), were included but not systematically. CONCLUSIONS: Research with more diverse faith leaders and that uses theory is needed to guide development of strategies for engaging this population in self-care to reduce obesity and related chronic diseases.

Activation of the Interleukin-18 Signaling Pathway via Direct Receptor Dimerization in the Absence of Interleukin-18.

Interleukin 18 (IL-18) is a key cytokine involved in the activation of T and NK cells, which are major effector cells in tumor killing. However, recombinant IL-18 showed limited efficacy in clinical trials. A recent study showed the lack of efficacy was largely due to the existence of IL-18BP, a soluble decoy receptor for IL-18. It was shown that engineered IL-18 variants that maintained pathway activation, but avoided IL-18BP binding, could exert potent antitumor effects. In this study, we demonstrated an alternative strategy to activate IL-18 signaling through direct receptor dimerization. These results provide evidences that the IL-18 pathway can be activated by directly bridging the receptors and, therefore, bypassing the IL-18BP-mediated inhibition.

Perceptions of mindfulness practices as a support for individuals managing caregiving responsibilities and chronic disease: A qualitative study.

OBJECTIVES: Explore the lived experience of individuals managing and/or caregiving for someone with a chronic disease and their perceptions of developing a mindfulness program for stress reduction. METHODS: Sixteen participants with chronic disease and/or caregivers participated. Participants completed eligibility screening, demographic questionnaires, and semi-structured interviews (30-60 min each) online or by phone. Interviews (n = 16) were audio recorded, transcribed, and analyzed using thematic analysis and NVivo® 12. Survey data were analyzed using SPSS® 28. RESULTS: Four themes emerged: (a) Chronic disease management and stress-perspectives on life's stressors; (b) Stress reduction techniques/perceptions of mindfulness-knowledge and implementation of stress reduction practices and familiarity with mindfulness; (c) Mindfulness program acceptability, barriers, and facilitators-interest, barriers, and facilitators to attending; (d) Mindfulness program structure-logistics to increase access and appeal to diverse audiences. DISCUSSION: Mindfulness has the potential for addressing the complexities of stress associated with disease management. Targeting mindfulness programs for populations with chronic disease management and caregiving responsibilities should include: Consideration of group formats with participation limited to this population, structuring programs to overcome barriers (i.e., culturally appropriate location), and equipping members of the community being served as instructors to ensure culturally relevant instruction.

Kinetics of sulfur-transfer from titanocene (poly)sulfides to sulfenyl chlorides: rapid metal-assisted concerted substitution.

The kinetics of sulfur transfer from titanocene (poly)sulfides (RCp2TiS5, Cp2TiS4CMe2, Cp2Ti(SAr)2, Cp2TiCl(SAr)) to sulfenyl chlorides (S2Cl2, RSCl) have been investigated by a combination of stopped-flow UV-Vis/NMR reaction monitoring, titration assays, numerical kinetic modelling and KS-DFT calculations. The reactions are rapid, proceeding to completion over timescales of milliseconds to minutes, via a sequence of two S-S bond-forming steps (k 1, k 2). The archetypical polysulfides Cp2TiS5 (1a) and Cp2TiS4C(Me2) (2a) react with disulfur dichloride (S2Cl2) through rate-limiting intermolecular S-S bond formation (k 1) followed by a rapid intramolecular cyclization (k 2, with k 2 ≫ k 1 [RSCl]). The monofunctional sulfenyl chlorides (RSCl) studied herein react in two intermolecular S-S bond forming steps proceeding at similar rates (k 1 ≈ k 2). Reactions of titanocene bisthiophenolates, Cp2Ti(SAr)2 (5), with both mono- and di-functional sulfenyl chlorides result in rapid accumulation of the monothiophenolate, Cp2TiCl(SAr) (6) (k 1 > k 2). Across the range of reactants studied, the rates are relatively insensitive to changes in temperature and in the electronics of the sulfenyl chloride, moderately sensitive to the electronics of the titanocene (poly)sulfide (ρ (Ti-(SAr)) ≈ -2.0), and highly sensitive to the solvent polarity, with non-polar solvents (CS2, CCl4) leading to the slowest rates. The combined sensitivities are the result of a concerted, polarized and late transition state for the rate-limiting S-S bond forming step, accompanied by a large entropic penalty. Each substitution step {[Ti]-SR' + Cl-SR → [Ti]-Cl + RS-SR'} proceeds via titanium-assisted Cl-S cleavage to generate a transient pentacoordinate complex, Cl-[Cp2TiX]-S(R')-SR, which then undergoes rapid Ti-S dissociation.

Associations Between Sociodemographic Variables, Social Determinants of Health, and Diabetes: Findings From a Congregational Health Needs Assessment.

PURPOSE: To examine associations between sociodemographic variables, social determinants of health (SDOHs) and diabetes using health needs assessment data. DESIGN: Cross-sectional study. SETTING: Faith-based communities in the Mid-South U.S. SAMPLE: Of the 378 churches, 92 participated in the study (24% response rate); N = 828 church leaders and members completed the survey. MEASURE: The Mid-South Congregational Health Survey assessed perceived health-related needs of congregations and the communities they serve. ANALYSIS: Generalized linear mixed modeling examined the associations between sociodemographic variables (age, sex, race/ethnicity, educational level), SDOHs (affordable healthcare, healthy food, employment), and diabetes. RESULTS: Individuals with less education had lower odds of reporting all SDOHs as health needs compared to individuals with more education (ORrange = .59-.63). Men had lower odds of reporting diabetes as a health need or concern compared to women (OR = .70; 95% CI = .50, .97). African Americans had greater odds of reporting diabetes as a health need compared to individuals in the 'Other' race/ethnicity category (OR = 3.91; 95% CI = 2.20, 6.94). Individuals who reported affordable healthcare (OR = 2.54; 95% CI = 1.73, 3.72), healthy food (OR = 2.24; 95% CI = 1.55, 3.24), and employment (OR = 3.33; 95% CI = 2.29, 4.84) as health needs had greater odds of reporting diabetes as a health need compared to those who did not report these SDOHs as needs. CONCLUSIONS: Future studies should evaluate strategies to merge healthcare and faith-based organizations' efforts to address SDOHs impacting diabetes.

The autophagy protein LC3A correlates with hypoxia and is a prognostic marker of patient survival in clear cell ovarian cancer.

Clear cell ovarian cancer histotypes exhibit metabolic features associated with resistance to hypoxia and glucose deprivation-induced cell death. This metabolic characteristic suggests that clear cell ovarian cancers activate survival mechanisms not typical of other epithelial ovarian cancers. Here we demonstrate that microtubule-associated protein 1 light chain 3A (LC3A), a marker of autophagy, is related to hypoxia and poor prognosis in clear cell ovarian cancer. In 485 ovarian tumours, we found that LC3A was significantly associated with poor progression-free (p = 0.0232), disease-specific (p = 0.0011) and overall patient survival (p = 0.0013) in clear cell ovarian cancer patients, but not in other subtypes examined. LC3A was an independent prognostic marker of reduced disease-specific [hazard ratio (HR): 2.55 (95% CI 1.21-5.37); p = 0.014] and overall survival [HR: 1.95 (95% CI 1.00-3.77); p = 0.049] in patients with clear cell ovarian carcinoma. We also found a strong link between autophagy and hypoxia as LC3A staining revealed a significant positive association with the hypoxia-related proteins carbonic anhydrase-IX and HIF-1α. The functional link between hypoxia and autophagy was demonstrated using clear cell and high-grade serous cell lines that were subjected to hypoxia or hypoxia + glucose deprivation. Clear cell carcinoma lines displayed greater autophagy induction and were subsequently more sensitive to inhibition of autophagy under hypoxia compared to the high-grade serous lines. Together, our findings indicate that hypoxia-induced autophagy may be crucial to the clinical pathology of clear cell ovarian cancer and is a potential explanation for histological subtype differences in patient disease progression and outcomes.

Chemical genetic strategy identifies histone deacetylase 1 (HDAC1) and HDAC2 as therapeutic targets in sickle cell disease.

The worldwide burden of sickle cell disease is enormous, with over 200,000 infants born with the disease each year in Africa alone. Induction of fetal hemoglobin is a validated strategy to improve symptoms and complications of this disease. The development of targeted therapies has been limited by the absence of discrete druggable targets. We developed a unique bead-based strategy for the identification of inducers of fetal hemoglobin transcripts in primary human erythroid cells. A small-molecule screen of bioactive compounds identified remarkable class-associated activity among histone deacetylase (HDAC) inhibitors. Using a chemical genetic strategy combining focused libraries of biased chemical probes and reverse genetics by RNA interference, we have identified HDAC1 and HDAC2 as molecular targets mediating fetal hemoglobin induction. Our findings suggest the potential of isoform-selective inhibitors of HDAC1 and HDAC2 for the treatment of sickle cell disease.

Functional screening identifies CRLF2 in precursor B-cell acute lymphoblastic leukemia.

The prognosis for adults with precursor B-cell acute lymphoblastic leukemia (B-ALL) remains poor, in part from a lack of therapeutic targets. We identified the type I cytokine receptor subunit CRLF2 in a functional screen for B-ALL-derived mRNA transcripts that can substitute for IL3 signaling. We demonstrate that CRLF2 is overexpressed in approximately 15% of adult and high-risk pediatric B-ALL that lack MLL, TCF3, TEL, and BCR/ABL rearrangements, but not in B-ALL with these rearrangements or other lymphoid malignancies. CRLF2 overexpression can result from translocation with the IGH locus or intrachromosomal deletion and is associated with poor outcome. CRLF2 overexpressing B-ALLs share a transcriptional signature that significantly overlaps with a BCR/ABL signature, and is enriched for genes involved in cytokine receptor and JAK-STAT signaling. In a subset of cases, CRLF2 harbors a Phe232Cys gain-of-function mutation that promotes constitutive dimerization and cytokine independent growth. A mutually exclusive subset harbors activating mutations in JAK2. In fact, all 22 B-ALLs with mutant JAK2 that we analyzed overexpress CRLF2, distinguishing CRLF2 as the key scaffold for mutant JAK2 signaling in B-ALL. Expression of WT CRLF2 with mutant JAK2 also promotes cytokine independent growth that, unlike CRLF2 Phe232Cys or ligand-induced signaling by WT CRLF2, is accompanied by JAK2 phosphorylation. Finally, cells dependent on CRLF2 signaling are sensitive to small molecule inhibitors of either JAKs or protein kinase C family kinases. Together, these findings implicate CRLF2 as an important factor in B-ALL with diagnostic, prognostic, and therapeutic implications.

Nickel-catalyzed ester carbonylation promoted by imidazole-derived carbenes and salts.

Millions of tons of acetyl derivatives such as acetic acid and acetic anhydride are produced each year. These building blocks of chemical industry are elaborated into esters, amides, and eventually polymer materials, pharmaceuticals, and other consumer products. Most acetyls are produced industrially using homogeneous precious metal catalysts, principally rhodium and iridium complexes. We report here that abundant nickel can be paired with imidazole-derived carbenes or the corresponding salts to catalyze methyl ester carbonylation with turnover frequency (TOF) exceeding 150 hour-1 and turnover number (TON) exceeding 1600, benchmarks that invite comparisons to state-of-the-art rhodium-based systems and considerably surpass known triphenylphosphine-based nickel catalysts, which operate with TOF ~7 hour-1 and TON ~100 under the same conditions.

A myeloid program associated with COVID-19 severity is decreased by therapeutic blockade of IL-6 signaling.

Altered myeloid inflammation and lymphopenia are hallmarks of severe infections. We identified the upregulated EN-RAGE gene program in airway and blood myeloid cells from patients with acute lung injury from SARS-CoV-2 or other causes across 7 cohorts. This program was associated with greater clinical severity and predicted future mechanical ventilation and death. EN-RAGEhi myeloid cells express features consistent with suppressor cell functionality, including low HLA-DR and high PD-L1. Sustained EN-RAGE program expression in airway and blood myeloid cells correlated with clinical severity and increasing expression of T cell dysfunction markers. IL-6 upregulated many EN-RAGE program genes in monocytes in vitro. IL-6 signaling blockade by tocilizumab in a placebo-controlled clinical trial led to rapid normalization of EN-RAGE and T cell gene expression. This identifies IL-6 as a key driver of myeloid dysregulation associated with worse clinical outcomes in COVID-19 patients and provides insights into shared pathophysiological mechanisms in non-COVID-19 ARDS.

Chemical phylogenetics of histone deacetylases.

The broad study of histone deacetylases in chemistry, biology and medicine relies on tool compounds to derive mechanistic insights. A phylogenetic analysis of class I and II histone deacetylases (HDACs) as targets of a comprehensive, structurally diverse panel of inhibitors revealed unexpected isoform selectivity even among compounds widely perceived as nonselective. The synthesis and study of a focused library of cinnamic hydroxamates allowed the identification of, to our knowledge, the first nonselective HDAC inhibitor. These data will guide a more informed use of HDAC inhibitors as chemical probes and therapeutic agents.

Examination of Associations between Physical Activity and Eating Identities among College-aged Adults Living in Hawai'i.

While physical activity and diet behaviors are correlated, mechanisms underlying associations have rarely been examined. This study examined associations between physical activity identity and eating identity among college-aged adults in Hawai'i to provide guidance for future multiple behavior change interventions. This study was a cross-sectional analysis of data collected between September 2013 and January 2014. Participants were 40 college students attending 4-year and 2-year institutions within the University of Hawai'i system. Total physical activity identity score and dimensions were measured using the Athlete Identity Questionnaire. Eating identity subtypes were measured using the Eating Identity Type Inventory. Associations between physical activity identity total score, 4 physical activity identity dimensions (appearance, importance, competence, and encouragement), and 4 eating identity subtypes (healthy, emotional, meat, and picky) were examined using multiple linear regressions. A significant positive association was found between total physical activity identity score and the healthy eating subtype and a negative association with the picky eating subtype. The physical activity dimension importance had a significant positive association with the healthy eating subtype, appearance a negative association with the emotional eating subtype, and competence a positive association with the meat eating subtype but a negative association with the picky eating subtype. The findings suggest important overlap in identities for physical activity and diet. Measurement of physical activity identity and eating identity as well as tailored intervention strategies should be incorporated into more behavior change research.

Congregational health needs by key demographic variables: Findings from a congregational health needs assessment tool.

Health needs assessments identify important issues to be addressed and assist organizations in prioritizing resources. Using data from the Mid-South Congregational Health Survey, top health needs (physical, mental, social determinants of health) were identified, and differences in needs by key demographic variables (age, sex, race/ethnicity, education) were examined. Church leaders and members (N = 828) from 92 churches reported anxiety/depression (65 %), hypertension/stroke (65 %), stress (62 %), affordable healthcare (60 %), and overweight/obesity (58 %) as the top health needs in their congregations. Compared to individuals < 55 years old and with a college degree, individuals ≥ 55 years old (ORrange=1.50-1.86) and with ≤ high school degree (ORrange=1.55-1.91) were more likely to report mental health needs (anxiety/depression; stress). African Americans were less likely to report physical health needs (hypertension/stroke; overweight/obesity) than individuals categorized as Another race/ethnicity (ORrange=0.38-0.60). Individuals with ≤ high school degree were more likely to report affordable healthcare as a need compared to individuals with some college or a college degree (ORrange=1.58). This research highlights the need for evaluators and planners to design programs that are comprehensive in their approach to addressing the health needs of congregations while also considering demographic variation that may impact program participation and engagement.

Tumor-infiltrating lymphocytes predict response to anthracycline-based chemotherapy in estrogen receptor-negative breast cancer.

INTRODUCTION: Infiltration of breast tumors by tumor-infiltrating lymphocytes (TIL) has been associated with sensitivity to anthracycline-based chemotherapy. However, it is unclear whether this is true within the estrogen receptor-alpha (ER)-negative subset of breast tumors that frequently manifest high TIL levels. METHODS: The association of TIL with short-term and long-term clinical response to anthracycline-based therapy was assessed in two independent ER-negative breast cancer cohorts in which patients were categorized as TIL-high or TIL-low. We defined an eight-gene lymphocyte mRNA expression signature (including CD19, CD3D, CD48, GZMB, LCK, MS4A1, PRF1, and SELL) and used unsupervised hierarchical clustering to examine the association between TIL and short-term response to neoadjuvant chemotherapy in a previously published cohort of ER-negative tumors (n = 113). We also examined the association between TIL and long-term chemotherapeutic efficacy in a second cohort of ER-negative tumors (n = 255) with longer than 6 years of median follow-up by using tissue microarrays and immunohistochemistry (IHC) for detection of CD3, CD8, CD4, CD20, and TIA-1. RESULTS: In patients with ER-negative tumors treated with neoadjuvant anthracycline-based chemotherapy, pathologic complete responses (pCRs) were achieved by 23 (74%) of 31 TIL-high patients and 25 (31%) of 80 TIL-low patients (odds ratio (OR), 6.33; 95% confidence interval (CI), 2.49 to 16.08; P < 0.0001). Multivariate logistic regression with standard clinicopathologic features demonstrated that only tumor size (P = 0.037) and TIL status (P = 0.001) were independent predictors of anthracycline response. In the second cohort, adjuvant anthracycline-based therapy was associated with increased disease-free survival (DFS) only in patients with high levels of intraepithelial CD3+ TIL (P = 0.0023). In contrast, outcomes after CMF treatment (cyclophosphamide, methotrexate, and fluorouracil) showed no association with CD3 status. In both cohorts, cytotoxic T-cells were the primary TIL subtype associated with anthracycline sensitivity. Finally, TIL significantly predicted anthracycline sensitivity for both the Her2-positive and triple-negative tumor phenotypes. CONCLUSIONS: ER-negative breast cancers with high levels of TIL have heightened sensitivity to anthracycline-based chemotherapy, as assessed by the immediate response to neoadjuvant therapy and long-term outcome following adjuvant therapy. Investigations of TIL-based predictive tests to identify patients likely to benefit from anthracycline-based treatments are warranted.