Hormone Response Research Articles

Abstract PO4-04-06: Exploring the use of tyrosine kinase inhibitors (TKIs) in the management of HER2-positive metastatic breast cancer post trastuzumab emtansine (T-DM1) therapy failure: Insights from a real-world study

Abstract Background Trastuzumab emtansine (T-DM1) is the established standard second-line treatment for HER2-positive metastatic breast cancer (MBC) patients who have previously received trastuzumab. However, there is ongoing debate regarding the subsequent therapeutic options following T-DM1 failure. This study aimed to evaluate the efficacy and safety of tyrosine kinase inhibitors (TKIs)-based therapy in HER2-positive MBC patients with resistance to T-DM1. Methods During the period from September 2018 to October 2021, a multicenter real-world study was conducted, enrolling a total of 53 patients diagnosed with HER2-positive metastatic breast cancer (MBC). These patients underwent TKIs-based therapy following the failure of T-DM1 treatment. The primary focus of the study was on evaluating progression-free survival (PFS) as the main endpoint. Secondary endpoints encompassed objective response rate (ORR), overall survival (OS), clinical benefit rate (CBR), and safety assessments. Results A total of 53 patients received TKIs-based therapy as a second-line or later treatment. The median progression-free survival (PFS) for TKIs-based therapy was 11.9 months (95% CI 7.9-15.9). The objective response rate (ORR) and clinical benefit rate (CBR) were found to be 19.6% and 72.5%, respectively. Among patients with brain metastasis (n=12), a median PFS of 10.5 months (95% CI 7.5-13.5) and an intracranial ORR of 33.3% were observed. Patients treated with pyrotinib (n=21) showed improved PFS compared to those treated with lapatinib (n=30), as well as patients who had derived favorable benefit from T-DM1 (PFS ≥ 6 months). Hormone receptor status (HR) and response to T-DM1 were identified as independent factors affecting the effectiveness of TKIs-based therapy. The most frequently reported adverse events included thrombocytopenia (23.6%), diarrhea (15.7%), leucopenia (15.7%), and hand-foot syndrome (15.7%). Conclusion TKIs-based therapy demonstrated favorable effectiveness and safety in HER2-positive metastatic breast cancer (MBC) patients who had previously received T-DM1 but experienced treatment failure. This includes patients with brain metastases. Notably, individuals who derived favorable benefits from prior T-DM1 treatment displayed significantly prolonged progression-free survival (PFS) when undergoing subsequent TKIs-based therapy. Citation Format: Yongmei Yin, Chunxiao Sun, Yijia Hua, Nan Jin, Wei Li, Xiang Huang. Exploring the use of tyrosine kinase inhibitors (TKIs) in the management of HER2-positive metastatic breast cancer post trastuzumab emtansine (T-DM1) therapy failure: Insights from a real-world study [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-04-06.

Abstract PO3-18-10: Impact of age and ovarian function suppression (OFS) on endocrine response to short preoperative endocrine therapy (ET): Results from the multicenter WSG-ADAPTcycle trial (n=5,290)

Abstract Background In HR+/HER2- early breast cancer (EBC), short preoperative endocrine therapy (ET) offers a promising tool for assessment of ET-efficacy based on Ki67-decrease after 2-4 weeks of treatment. Low post-endocrine Ki67 (Ki67post) is associated with good prognosis in several large prospective trials. WSG-ADAPT demonstrated that Ki67post is a promising tool for decision making in cases with uncertain adjuvant chemotherapy indication, e.g., in premenopausal patients (pts.) with N0 and Recurrence Score (RS, Oncotype DX®) RS 16-25 or N1 and RS ≤25. Preliminary results from the phase III ADAPTcycle trial indicated higher efficacy of preoperative ET in premenopausal pts. if ovarian function suppression (OFS) was used together with tamoxifen (TAM) or aromatase inhibitors (AI). In the ADAPTcycle screening population, we are now able to validate these results and investigate the influence of age subgroups, Recurrence Score, and individual biological markers, as well as OFS on ET-response. Methods In ADAPTcycle (n=5,290 screened until 06/23 at 84 sites in Germany), N0-1 pts. with RS >25 or N2-3 pts with RS ≤25 and ET-response (Ki67post ≤10%) were randomized to (neo)adjuvant standard chemotherapy (CT) or ribociclib + AI +/- GnRH-analog (n=1,670 randomized). Participation of premenopausal pts. with N1-disease and RS ≤25 or N0 and RS 16-25 was allowed irrespective of ET-response, but randomization recommended only for ET-responders. Use of OFS + TAM or AI for ET response assessment in the preoperative phase was protocol-recommended. This analysis includes all patients with baseline RS and data on ET-response. Ki67-response is defined as Ki67post ≤10% (central pathology assessment) after 2-4 weeks of therapy (OFS-use recommended for 4 weeks). ER-, PR-, and HER2-levels are analyzed by IHC and mRNA. Results Results on ET-response according to age and clinicopathological as well as tumor biological characteristics will be presented at the meeting. So far, ADAPTcycle with >5,000 pts. screened is the largest trial worldwide to look at ET response assessment in pre- and postmenopausal pts. with HR+/HER2- EBC. The results thus have the potential to impact clinical practice. Contact Information oleg.gluz@wsg-online.com Citation Format: Oleg Gluz, Matthias Christgen, Ulrike Nitz, Sherko Küemmel, Michael Braun, Marc Thill, Rachel Wuerstlein, Pauline Wimberger, Andreas Hartkopf, Christian Schem, Matthias Zaiss, Vesna Bjelic-Radisic, Marianne Just, Kristina Veselinovic, Myriam Vincent, Monika Graeser, Katja Krauss, Oliver Hoffmann, Kerstin Lüdtke-Heckenkamp, Ronald Kates, Christine zu Eulenburg, Peter Schmid, Rick Baehner, Hans-Heinrich Kreipe, Nadia Harbeck. Impact of age and ovarian function suppression (OFS) on endocrine response to short preoperative endocrine therapy (ET): Results from the multicenter WSG-ADAPTcycle trial (n=5,290) [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO3-18-10.

Abstract PO2-19-07: PROOFS – Pre-/perimenopausal patients with HR+/HER2- EBC with intermediate to high clinical and low genomic risk, treated by endocrine treatment (ET) plus ovarian function suppression (OFS) or chemotherapy followed by ET

Abstract Background During the past years, clinical routine has shifted away from relying solely on clinicopathologic factors toward increasing use of multigene expression assays in guiding treatment decisions regarding adjuvant chemotherapy (CT) for hormone receptor-positive (HR+), HER2-negative (HER2-) early breast cancer (EBC) with 0 to 3 positive lymph nodes. In case of a low genomic risk, there is strong consensus in favor of endocrine treatment (ET) alone for postmenopausal patients, regardless of clinical risk (supported by data from planB, ADAPT, RxPonder, TailorX, MINDACT). However, there is still uncertainty regarding optimal therapy for pre- and perimenopausal patients with a similar clinical/genomic risk profile. Since the observed benefit of CT in this patient group may be partly attributable to CT-induced ovarian function suppression (OFS), some of these patients could be spared CT by adding OFS to endocrine therapy. The West German Study Group (WSG) initiated the PROOFS-registry to create a real-world database on how to optimally treat these patients. Eligibility Criteria PROOFS (NCT05792150) aims at the long-term follow up of pre- or perimenopausal patients with HR+/HER2- EBC with an intermediate to high clinical risk of recurrence and low genomic risk measured by MammaPrint(R). Additional endocrine response assessment (Ki67) after a short preoperative endocrine therapy is encouraged. Up to 25% of the study population may have node-positive disease. Participants undergo standard-of-care treatment at physician´s discretion and according to clinical routine, which may include CT neoadjuvant or adjuvant + ET, ET + OFS in premenopausal, or ET +/- OFS in perimenopausal women. Specific Aims PROOFS aims to give insights in the real-world use of OFS and to confirm an excellent outcome in pre-/perimenopausal women with HR+/HER2- early BC with an intermediate to high clinical risk of recurrence and low genomic risk treated by ET +/- OFS alone (without CT). In addition, quality of life is captured. Statistical Methods The primary objective is to demonstrate the 5-year distant recurrence-free interval (dRFI) in patients treated by ET +/- OFS alone. The null hypothesis H(0) is: 5-year dRFI < 92%. Secondary endpoints for all patients are 5- and 10-year dRFI, distant disease-free survival (dDFS), overall survival (OS), breast cancer-free interval (BCFI), quality of life (QLQ BR23 and QLQ-C30) and therapy adherence. Cox regression models, Kaplan-Meier method and log-rank tests will be applied for survival analysis. Linear mixed models will be utilized to quantitatively describe the course of quality of life-scores as well as therapy adherence, and to conduct group comparisons. Present and Target Accrual The study is open for recruitment of 1500 patients (approximately 20% receiving CT and 80% receiving ET +/- OFS) at up to 100 centers in Germany (June 2023: 24 open for recruitment). Funding AGENDIA N.V., Amsterdam Contact Information rachel.wuerstlein@med.uni-muenchen.de Citation Format: Rachel Wuerstlein, Lotta Ada Fischer, Nadia Harbeck, Monika Graeser, Ulrike Nitz, Matthias Christgen, Sherko Kuemmel, Christine zu Eulenburg, Matthias Warm, Oleg Gluz. PROOFS – Pre-/perimenopausal patients with HR+/HER2- EBC with intermediate to high clinical and low genomic risk, treated by endocrine treatment (ET) plus ovarian function suppression (OFS) or chemotherapy followed by ET [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-19-07.

Abstract PO2-01-08: Differential HOXB13 gene expression and promoter methylation analysis in breast cancer

Abstract Background: The Breast Cancer Index (BCI) is a gene expression-based signature comprising two functional biomarker panels: the Molecular Grade Index (MGI) and BCI (H/I), which is a ratio of the HOXB13 and IL17BR gene expression. Several studies have shown that BCI (H/I) is a predictive biomarker for extended adjuvant endocrine therapy benefit in hormone receptor-positive (HR+) early-stage breast cancer. Molecular mechanisms underlying endocrine responsiveness remain to be fully explored. The objective of this analysis is to evaluate the correlation between gene expression and methylation in breast cancer patients displaying different HOXB13 mRNA expression levels from The Cancer Genome Atlas (TCGA) project. Methods: Data from methylation microarrays and mRNA sequencing were examined in combination with clinical metadata from 1,095 TCGA breast cancer patients. After removing outliers, HR+ samples were ordered based on their HOXB13 expression levels and the top 10% of samples (HOXB13-high; n=95) and bottom 10% (HOXB13-low; n=95) were selected for further analysis. Differentially expressed genes (DEGs) were determined by comparing all protein-coding genes between the HOXB13-high and HOXB13-low sets, based on an absolute log-fold change above 2 and a Benjamini-Hochberg adjusted p-value threshold of 0.01. Among the subset of samples with matching methylation data (n=60 for the HOXB13-high group and n=48 for the HOXB13-low group), hypo-methylated and hyper-methylated differentially methylated probes (DMPs) were identified based on at least 10% change of β (i.e., average promoter methylation) and a Benjamini-Hochberg adjusted p-value threshold of 0.01. Motif and transcription factor (TF) enrichment analyses were identified at a minimum incidence of 10 and a lower boundary of 1.5 for the 95% confidence interval of the odds ratio. Results: Our analysis identified a total of 613 DEGs between the HOXB13-high and HOXB13-low groups. Gene ontology analysis of DEGs revealed a statistically significant enrichment of genes associated with PTK6 regulated cell cycle, oncogene-induced cellular senescence, and regulation of T cell activation. Analysis of protein-protein interactions of differentially expressed genes revealed differences in key biological processes including changes in cell cycle regulation, p53 pathway, and IL-17 signaling. A total of 5,295 hyper-methylated and 580 hypo-methylated DMPs were discovered when comparing HOXB13-high and HOXB13-low samples. Changes in global methylation patterns enabled classification of breast cancer samples with high or low HOXB13 expression with an accuracy of 0.90. Reconstruction of gene regulatory networks from DNA methylation and transcriptome profiles elucidated significant pairs of DMPs and differentially expressed genes. Specifically, a total of 197 differentially expressed genes were attributed to hyper-methylated DMPs, including genes involved in response to estrogen, activation of HOX genes during differentiation, and drug-mediated inhibition of CDK4/6 activity. Additionally, 8 differentially expressed genes were attributed to hypo-methylated DMPs and were associated with proteasomal protein catabolic processes. Lastly, motif analysis of transcriptional factors for the hyper-methylated DMPs revealed enrichment of binding motifs attributed to FOXA1, ESR1, XBR1, and FOXP1. Conclusion: A comprehensive and comparative analysis of mRNA expression levels and methylation patterns between TCGA breast cancer samples with high and low expression of HOXB13 revealed key signaling pathways and biological processes that may provide insights on the molecular mechanism of HOXB13 in the regulation of response to endocrine therapy in HR+ breast cancer. Citation Format: Natalia Siuliukina, Yi Zhang, Kai Treuner, Mark Pegram. Differential HOXB13 gene expression and promoter methylation analysis in breast cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-01-08.

Abstract LBO1-05: Impact of age and ovarian function suppression (OFS) on endocrine response to short preoperative endocrine therapy (ET): Results from the multicenter ADAPTcycle trial (n=4,334)

Abstract Background: In HR+/HER2- early breast cancer (EBC), short preoperative endocrine therapy (ET) is a promising tool for ET-efficacy assessment based on Ki67-decrease after 2-4 weeks of ET. Low post-endocrine Ki67 (Ki67post) is associated with good prognosis in large prospective trials. WSG-ADAPT demonstrated that ET-response is a valid criterion for decisions in uncertain adjuvant chemotherapy indications, e.g., in premenopausal patients (pts) with N0 and Recurrence Score (RS, Oncotype DX®) RS 16-25 or with N1 and RS ≤25. Preliminary results from the phase III ADAPTcycle trial indicated higher efficacy of preoperative ET in premenopausal pts. if ovarian function suppression (OFS) was used together with tamoxifen (TAM) or aromatase inhibitors (AI). In the final screening population, we are now able to confirm these results and investigate the influence of age subgroups, RS, individual biological markers, and OFS on ET-response. Methods: In ADAPTcycle (n=5,290 screened, 1,670 randomized by 06/23; 84 sites in Germany), N0-1 pts with RS >25 or N2-3 pts with RS ≤25 and ET-response were randomized to (neo)adjuvant chemotherapy (CT) followed by ET vs. ribociclib + AI (premenopausal: +GnRH). In premenopausal pts, participation with N1-disease and RS ≤25 or N0 and RS 16-25 was allowed irrespective of ET-response, but randomization recommended only for ET-responders; in the preoperative phase, OFS+TAM or AI for ET-response assessment was protocol-recommended. Results: This analysis includes all pts with complete baseline characteristics (including RS) and ET-response data (n=4,334); ET-response was defined as Ki67post ≤10% (central pathology) after 2-4 weeks (if OFS: 4 weeks) of therapy. ER, PR, and HER2-levels were analyzed by IHC and mRNA. Median age was 56 years (22-87y). 55% had cT2-4 tumors; 28% had cN+ disease; 42% were G3 (central lab). Median RS was 20, median baseline Ki67 25%. n=1,368 were ≤50y and premenopausal (“≤50y”): n=314 (23.0%) and 255 (18.6%) were treated by AI+OFS and TAM+OFS, respectively. 2,966 were >50y or postmenopausal (“ >50y”), of whom 2,565 received AI (86.5%). ET-response occurred in 48.2% of “≤50y” vs. 72.7% in “ >50y” pts (p< 0.001). We observed significant differences in ET-response rates between “≤50y” vs. “ >50y” with TAM alone (34.7% vs. 46.4%); In “≤50y” pts, 55.7% ET-response occurred with TAM+OFS vs. 76.4% with AI+OFS, comparable to 76.8% in “ >50y” patients with AI. Moreover, OFS led to higher ET-response rates (by uni- and multivariable analysis) even in the small (n=78) group of premenopausal pts >50y. Remarkably, even in pts with RS >25, substantial ET-response rates were observed with AI: 66.7% (“≤50y”, AI+OFS), 55.6% (“ >50y”, AI) vs. lower rates with TAM alone (>50y: 18.2%; < 50y: 15.8%). ET-response rate in “≤50y” pts was associated with ET type, RS, and baseline Ki67 in both uni- and multivariable analysis, but not age subgroup (≤40 vs. 41-50y). ER, PR, and HER2 expression by IHC or RT-PCR were associated with ET-response by univariable (but not multivariable) analysis. In “ >50y” pts, by uni- and multivariable analysis, again ET-type, RS, and baseline Ki67, as well as ER-expression (by IHC) and ESR1, PR, and HER2-expression (by RT-PCR) were associated with higher ET-response rates. Conclusions: With about 10,000 pre- and postmenopausal pts, ADAPT and ADAPTcycle provide the largest international database regarding ET-response. ADAPTcycle confirms ADAPT ET-response rates in pts receiving TAM or AI; adding OFS to either improves ET-response, irrespective of RS. ET-response measurement adds value in therapy decisions, particularly in ≤50y N0-1 pts. ET-response might provide key information even in pts with high RS; outcome data of ADAPTcycle will reveal potential clinical consequences. Optimal preoperative ET, especially in premenopausal pts, might even enable sparing chemotherapy in additional pts. Citation Format: Oleg Gluz, Matthias Christgen, Ulrike Nitz, Sherko Küemmel, Michael Braun, Marc Thill, Rachel Wuerstlein, Pauline Wimberger, Andreas Hartkopf, Christian Schem, Matthias Zaiss, Vesna Bjelic-Radisic, Marianne Just, Kristina Veselinovic, Myriam Vincent, Monika Graeser, Katja Krauss, Oliver Hoffmann, Kerstin Lüdtke-Heckenkamp, Ronald Kates, Christine zu Eulenburg, Katarzyna Jozwiak, Sandy Burmeister, Rick Baehner, Peter Schmid, Hans-Heinrich Kreipe, Nadia Harbeck. Impact of age and ovarian function suppression (OFS) on endocrine response to short preoperative endocrine therapy (ET): Results from the multicenter ADAPTcycle trial (n=4,334) [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr LBO1-05.

Abstract PO5-08-07: Ki-67 Expression in Breast Cancer associated with ATM, BRCA1, BRCA2, CHEK2 and PALB2 Pathogenic Variants

Abstract Background: Germline Pathogenic Variants (PVs) in ATM, BRCA1, BRCA2, CHEK2, and PALB2 are known to increase breast cancer (BCa) risk, but the effect of these PVs on tumor biology is not well-understood. Ki-67 is an established prognostic marker in hormone receptor (HR) positive breast cancer but the distribution of Ki-67 in breast cancer associated with germline PVs is not known. Methods: The study included patients with loco-regional (Stage I-III) HR-positive (ER+ or PR+), HER2-negative invasive ductal or lobular breast cancer evaluated at Mayo Clinic for curative intent treatment between 2012 and 2021. PV carrier status was ascertained in more than two-thirds of cases from germline whole exome sequencing (Regeneron) of patients who participated in a prospective registry at Mayo Clinic. Additional PV carriers were identified through a review of the results of clinical germline genetic testing in electronic medical records aided by natural language processing. Clinically performed Ki-67 values were abstracted from electronic medical records and analyzed as continuous and categorical variables (low: < 20% vs. high: ≥20%). Throughout the timeline of the study, a consistent method for assessment of Ki-67 was utilized at Mayo Clinic, Rochester: Immunohistochemical staining of the Ki-67 antigen in formalin-fixed paraffin-embedded tissue was performed using MIB-1 clone, the scanned slides were reviewed by a technologist and traced areas further analyzed for Ki-67 using Aperio Software. Patients who received neoadjuvant therapy prior to the assessment of Ki-67, with metastatic disease at diagnosis, with PVs in two or more genes of interest, or unknown germline PV or Ki-67 status were excluded from the analysis. Multiple primary tumors in the same patient diagnosed within one year of each other were analyzed independently. The control (non-carrier) group included randomly selected patients from the prospective registry who tested negative for PVs in the five genes over the same time period. Ki-67 values were compared between germline PV carriers and non-carriers in a multivariable logistic regression analysis adjusting for sex, race, age at diagnosis, stage at presentation, and histology of the tumor. Results: A total of 113 HR+/HER2- tumors from 111 PV carriers (ATM: 27, BRCA1: 24, BRCA2: 28, CHEK2: 22, and PALB2:12) along with 431 HR+/HER2- tumors from 422 randomly selected non-carriers were evaluated. The majority (>95%) of patients were non-Hispanic White. Germline PV carriers in each gene were diagnosed at a significantly younger age (P< 0.05) compared to non-carriers. The median Ki-67 value was 12 (Range: 1 to 93) for non-carriers, 16 (Range: 0 to 79) for ATM, 31 (Range: 4 to 86) for BRCA1, 30 (Range: 6 to 80) for BRCA2, 13 (Range: 1 to 38) for CHEK2, and 20 (Range: 3 to 46) for PALB2 PV carriers. In multivariable analysis, compared to non-carriers, we observed significantly higher odds of a high Ki67 value (≥20%) for tumors arising from germline PV carriers in BRCA1 (Odds Ratio (OR): 7.3, 95%CI: 2.3 – 23.3, p< 0.001), BRCA2 (OR: 3.0, 95%CI: 1.2 – 7.4, p=0.02) and PALB2 (OR: 3.9, 95%CI: 1.1 – 14.5, p=0.04). No such relationship was observed with either ATM (OR: 1.1, 95%CI: 0.4 – 2.9, p=0.81) or CHEK2 (OR: 0.8, 95%CI: 0.3 – 2.2, p=0.63). Conclusion: This study reveals that HR+/HER2- tumors in BRCA1, BRCA2, and PALB2 PV carriers have a significantly higher Ki-67 expression compared to non-carriers. These findings have implications for understanding prognosis and endocrine response PV with HR+ breast cancer. Further exploration of the relationships between Ki67 and germline PV status is necessary to personalize systemic therapy options in germline PV carriers. Acknowledgments: Thank you to Regeneron Genetics Center. The study was funded in part by the NIH Specialized Program of Research Excellence in Breast Cancer [P50CA116201] to Mayo Clinic, Mayo Clinic Department of Oncology Small Grants Program, and the Paul Calabresi Program at Mayo Clinic Citation Format: Robert Scheel, Grace Choong, Tara Rao, Nicholas Boddicker, Chunling Hu, Jie Na, Vinod Kaggal, Sean Murphy, Karthik Giridhar, Saba Yasir, Matthew Goetz, Fergus Couch, Siddhartha Yadav. Ki-67 Expression in Breast Cancer associated with ATM, BRCA1, BRCA2, CHEK2 and PALB2 Pathogenic Variants [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-08-07.

Abstract PO4-05-14: Gene expression in metastatic breast cancer – patterns and prognostic potential

Abstract Introduction Metastatic breast cancer (mBC) is the leading cause of breast cancer mortality. However, mBC has a considerable variance in prognosis and course of disease, reflecting a substantial biological heterogeneity. Despite this, there is a lack of prognostic biomarkers validated for mBC. New prognostic markers and improved understanding of the molecular variants of mBC could aid in prognostication and personalizing treatments, and lead to the discovery of novel drug targets. The primary objective of this study was to explore the prognostic value of gene expression (GEX) in the distant metastasis (DM) of mBC. Secondly, we aimed to identify molecular subtypes of mBC, and describe their biological niches. Methods We used a cohort of mBC patients enrolled in a prospective trial (NCT01322893) that includes tissue from primary tumor (PT) and matched lymph node metastasis (LNM) and/or DM (1). The inclusion criteria were diagnosed mBC with a life expectancy of >2 months, ECOG score of 0-2, and an age of >18 years. RNA was extracted from formalin-fixed, paraffin-embedded tumor tissue. GEX data were acquired for n=123 PT, n=71 LNM, and n=74 DM using the NanoString Breast Cancer 360 assay, which comprises 758 genes and >30 GEX signatures. The relationships between GEX and the endpoints overall survival (OS) and progression-free survival (PFS) were assessed using cox proportional hazards regression with GEX as a continuous score. Multivariable cox regressions were adjusted for PAM50 of the DM, number of metastatic sites, visceral metastasis, ECOG, and age at mBC diagnosis. Visualization by heatmap and k-means clustering were used to identify GEX patterns. When clustering single genes, we included only the 100 genes with the highest standard deviation. Results In relation to PFS, we found prognostically unfavorable roles of DM GEX of the signatures for Treg cells (HR=1.3, p=0.02), cytotoxicity (HR=1.3, p=0.03), and p53 (HR=1.3, p=0.02). Further, our data indicate prognostically favorable value of DM GEX of ESR1 (HR=0.77, p=0.02), PGR (HR=0.65, p=0.002), FOXA1 (HR=0.76, p=0.02), and AR (HR=0.68, p=0.0006). The prognostic value of AR remained in multivariable analysis. In relation to OS the first 2 years after mBC prognosis, we found prognostically favorable value of DM GEX of B7H3 (HR=0.65, p=0.04), TGFB (HR=0.53, p=0.02) and the signatures for stroma (HR=0.7, p=0.05) and claudin-low (HR=0.61, p=0.02). Finally, our data suggests that the previously published PAM50MET panel performs well as a prognostic indicator, especially when considered on a continuous scale, for PFS (HR=1.3, p=0.0023) and OS (HR=1.3, p=0.003) (2). Based on the 100 most highly variable genes, we identified five stable tumor clusters with distinct GEX profiles, where some strongly associated with metastatic site. Using the same approach on the GEX signatures, we established four GEX profiles able to outperform PAM50 in identifying tumors with poor OS in this material. Conclusion In this study, we demonstrate prognostic value for GEX-based panels in DM samples. First, we find GEX signatures related to hormone responsiveness to be favorable, and genetic instability to be unfavorable for mBC prognosis. We also demonstrate AR to be an independent marker for better PFS. Further, we confirm the prognostic value of PAM50MET, a previously published panel with the advantage of including only PAM50 genes and clinical parameters. Finally, we identify new GEX profiles related to metastatic site and outcome. These results illuminate the biological differences between mBC in relation to outcome and metastatic site. Better understanding of the mBC GEX subtypes may open new venues for tailored treatment.

Male breast cancer differs from female breast cancer in molecular features that affect prognoses and drug responses

BackgroundMale breast cancer (MBC) is a rare malignancy with a worse prognosis than female breast cancer (FBC). Current MBC treatment strategies are based on those for FBC. However, molecular differences between MBC and FBC with respect to prognosis and drug responses remain unclear. MethodsAfter controlling for confounding factors with propensity score matching (PSM), differences between MBC and FBC were comprehensively analyzed using many types of data: survival, immune microenvironments, sex hormone responses, drug sensitivity, transcriptomes, genomes, epigenomes, and proteomes. ResultsOverall survival (OS) and cancer-specific survival (CSS) were both worse for MBC than for FBC. Differentially expressed mRNAs were enriched in numerous cancer-related functions and pathways, with SPAG16 and STOX1 being as the most important prognosis-related mRNAs for MBC. Competing endogenous RNA (ceRNA) and transcription factor (TF)-mRNA regulatory networks contain potential prognostic genes. Nine genes had higher mutation frequencies in MBC than in FBC. MBC shows a comparatively poor response to immunotherapy, with five proteins that promote breast cancer progression being highly expressed in MBC. MBC may be more responsive than FBC to estrogen. We detected six United States Food and Drug Administration (FDA)-approved therapeutic target genes as being differentially expressed between MBC and FBC. ConclusionThe poor prognosis of MBC compared to FBC is due to numerous molecular differences and resulting drug responses.

Resistance Exercise Sessions Comprising Multijoint vs. Single-Joint Exercises Result in Similar Metabolic and Hormonal Responses, But Distinct Levels of Muscle Damage in Trained Men.

Barbosa, PH, Bueno de Camargo, JB, Jonas de Oliveira, J, Reis Barbosa, CG, Santos da Silva, A, Dos-Santos, JW, Verlengia, R, Barreira, J, Braz, TV, and Lopes, CR. Resistance exercise sessions comprising multijoint vs. single-joint exercises result in similar metabolic and hormonal responses, but distinct levels of muscle damage in trained men. J Strength Cond Res 38(5): 842-847, 2024-Resistance-type exercise (RE) elicits distinct acute metabolic and hormonal responses, which can be modulated by the manipulation of training variables. The purpose of this study was to compare the metabolic (blood lactate and estimated lactic anaerobic system energy expenditure) and hormonal (growth hormone [GH]) responses to RE sessions composed exclusively of multijoint (MULTI) or single-joint (SINGLE) exercises. Assessments of creatine kinase (CK) levels were also performed. In a crossover design, 10 recreationally resistance-trained men (age: 26.9 ± 3.0 years, total body mass: 83.2 ± 13.8 kg; height: 176 ± 7.0 cm; training experience: 5.5 ± 2.4 years) were randomly submitted to both protocols. Blood collections were made pre, 3 minutes after, and 36 hours after each experimental session. No significant difference between MULTI vs. SINGLE was observed for the rises in blood lactate (p = 0.057) and GH (p = 0.285) levels. For CK, a significant difference between the protocols was noted, in which MULTI resulted in significant rises after 3 minutes (p = 0.017) and 36 hours (p = 0.043) compared with SINGLE. In conclusion, the findings of this study suggest that resistance-trained individuals display similar metabolic and hormonal responses when performing MULTI and SINGLE exercise protocols. Also, RE sessions comprising MULTI exercises induce a higher magnitude of muscle damage, which may require a longer recovery period compared with SINGLE.

The effect of methylphenidate on the dopamine and growth hormone response to exercise in children with attention-deficit hyperactivity disorder

PurposeTo assess the growth hormone (GH) and Dopamine (DA) response to exercise in children with attention-deficit hyperactivity disorder (ADHD) with and without methylphenidate (MP). We hypothesized that the GH and DA response to the exercise with MP would be siginicantly lower. MethodsTwenty children participated in the study (12 males and 8 females, age range 9–13 years). Ten with ADHD and 10 controls. Participants with ADHD performed an exercise test twice, with and without MP while controls performed one exercise test. Blood samples for GH and DA were collected before, at peak, 30 and 60 min after the end of exercise. ResultsCompared to controls, children with ADHD with and without MP, had a significantly lower GH (P < .002) and DA (P < .01) responses to exercise. In participants with ADHD, a significantly greater GH response (p < .04) to exercise was found when MP administered to the children before exercise, yet this response was still significantly lower than controls. ConclusionsGH and DA excretion after an exercise challenge in children with ADHD is impaired. MP slightly attenuates the GH blunted response. This may link ADHD with growth impairment in some children and explain previous findings indicating that the final adult height is usually not compromised in children with ADHD treated with MP. The combined exercise and stimulant treatment therapeutic effects needs to be further explored.Trial registration number: NCT00945971

Effect of estrogens on apoptosis in the pituitary of viscachas (Lagostomus maximus maximus)

Estrogens, acting through their receptors (ERα and ERβ), regulate cell turnover in the pituitary gland, influencing cell proliferation and apoptosis across various species. However, their role in pituitary processes in seasonally reproducing animals remains poorly understood. This study aims to investigate the influence of estrogens, through the expression of their specific receptors, on the apoptosis of PD cells in relation to sexual maturity, the reproductive cycle, and pregnancy in a seasonal reproductive rodent (Lagostomus maximus maximus). ERα and caspase-3-cleaved (CASP3c) immunoreactive (-ir) cells were identified through immunohistochemistry. Apoptotic cells were detected using the TUNEL technique, with quantitative analysis facilitated by image analysis software, alongside measurement of serum estradiol levels using radioimmunoassay The immunostaining pattern for ERα included nuclear (ERαn) and cytoplasmic (ERαc) staining. In male viscachas, ERα expression significantly increases from immature to adult animals, correlating with the rise in serum estradiol levels and a decrease in the percentage of apoptotic cells. During the gonadal regression period in adult males, a decrease in the number of ER-ir cells and serum levels of estradiol corresponds with an increase in the number of apoptotic cells. In females, serum levels of estradiol peaked during mid-pregnancy, coinciding with a significant decrease in the number of apoptotic cells in the PD. Simultaneously, the percentage of ERαn-ir cells reaches its maximum value during late pregnancy, indicating the need to maintain the protective action of this gonadal hormone throughout the extensive pregnancy in these rodents. Regional ERα receptor expression and apoptotic cells appear to be associated with distinct PD cell populations and their hormonal responses. Finally, elevated estradiol levels coincide with diminished apoptotic cells in the male reproductive cycle and during pregnancy, suggesting an antiapoptotic role of estradiol in this species.

Prefrontal-medullary Circuitry Impacts Sex-specific Microvascular Function Following Chronic Stress

Vascular dysfunction is a prominent factor underlying cardiovascular disease, the leading cause of death worldwide. Additionally, chronic stress contributes to cardiovascular pathologies in a sex-dependent manner. However, the neurobiological mechanisms mediating the sex-specific impact of chronic stress on vascular function are not known. Our previous studies found that the prefrontal infralimbic cortex (IL)-rostral ventrolateral medulla (RVLM) circuit inhibits sympathetic and endocrine responses to stress; therefore, the current study tested the hypothesis that the IL-RVLM circuit is necessary to mitigate microvascular dysfunction following chronic stress. To this end, an intersectional genetic approach was used to induce Cre-dependent expression of tetanus toxin light chain and inhibit neurotransmitter release from RVLM-projecting IL neurons in male and female rats. Rats were then exposed to 2 weeks of chronic variable stress (CVS) or remained unstressed. Left ventricle structure was assessed by echocardiography and microvascular function was subsequently assessed by resistance arteriole pressure myography. Arteriole myogenic tone, endothelial-independent vasoconstriction, as well as endothelial-dependent vasodilation and vasoconstriction were determined at both normotensive (70 mmHg) and hypertensive (120 mmHg) pressures. Our results indicate that heart size and ventricular thickness were impacted in a sex-dependent manner. Circuit inhibition increased heart index in females regardless of stress. In males, circuit inhibition reduced ventricular wall thickness in diastole. Microvasculature also exhibited sex-dependent function. Following CVS, males had increased myogenic tone compared to females. Circuit inhibition also differentially impacted vasodilation in males and females. In chronically stressed females, circuit inhibition increased endothelial-dependent vasodilation at both normotensive and hypertensive pressures and normotensive endothelial-independent relaxation. These changes indicate that chronic stress leads to female microvascular dysfunction that is exacerbated by IL-RVLM inhibition. Chronically stressed males had no changes in normotensive vasodilation. However, with elevated pressure, circuit-inhibited CVS males had increased endothelial-dependent vasodilation. In contrast, unstressed males with circuit inhibition had decreased endothelial-dependent vasodilation at 120 mmHg, indicating that the male IL-RVLM circuit may be necessary for vascular function during acute hypertensive events. Broadly, these results support the hypothesis that the IL-RVLM circuit is necessary for appropriate vascular function and that circuit inhibition and chronic stress lead to sex-specific responses that may differentially impact the progression of cardiovascular disease. NIH R01 HL150559 (B.Myers) NIH F30 OD032120 (C. Dearing). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Immunohistochemical Characterization of Breast Carcinoma: Clinical Correlations, Molecular Subtypes, and Therapeutic Implications

Background: Breast carcinoma exhibits heterogeneity in terms of morphology, molecular, treatment response, and clinical outcomes. The objective of the study was to classify the various malignant breast cancer cases based on their immunohistochemical characteristics and understand their association and behavior, which may be useful for predicting treatment and prognosis. Methods: In this study, 12808 malignant breast cancer cases were studied based on hormone receptor IHC biomarkers, age, gender, histological type, grade and molecular classifiers. Results: The mean age of patients was 53.63+13.08 years, around 45.43% were grade 3 tumors and the invasive duct carcinoma of non-specified type (NST) was the most common type seen (93.83%). ER positivity was 83.89% in grade 1, 69.9% in grade 2, and 36.86% in grade 3 tumors, and the overall PR and Her-2 positivity was 47.06% and 18.67%, respectively. A relatively higher percentage of Triple Negative cases was seen (36.58%), followed by 25.43% Luminal A cases. There was a significant association between molecular subtypes with respect to age, gender, Scarff Bloom Richardson (SBR) grade, and histological type (P&lt;0.0001). Overall, grade 3 tumor cases were most common, the majority of which were Triple Negative. Maximum cases of triple negative tumors were seen among women, being mostly concentrated in younger age group i.e. &lt;40 years. Conclusion: Immunohistochemistry for hormone receptor positivity remains the mainstay of diagnosis and molecular sub-classification. The hormone response of tumors is important prognostically, in predicting the treatment outcomes. Going further, molecular analysis and gene expression studies can further augment the histopathological diagnosis to assist strategies of targeted therapy and precision medicine resulting in better patient outcomes.

Identification and characterization of ACR gene family in maize for salt stress tolerance.

Members of the ACR gene family are commonly involved in various physiological processes, including amino acid metabolism and stress responses. In recent decades, significant progress has been made in the study of ACR genes in plants. However, little is known about their characteristics and function in maize. In this study, ACR genes were identified from the maize genome, and their molecular characteristics, gene structure, gene evolution, gene collinearity analysis, cis-acting elements were analyzed. qRT-PCR technology was used to verify the expression patterns of the ZmACR gene family in different tissues under salt stress. In addition, Ectopic expression technique of ZmACR5 in Arabidopsis thaliana was utilized to identify its role in response to salt stress. A total of 28 ZmACR genes were identified, and their molecular characteristics were extensively described. Two gene pairs arising from segmented replication events were detected in maize, and 18 collinear gene pairs were detected between maize and 3 other species. Through phylogenetic analysis, three subgroups were revealed, demonstrating distinct divergence between monocotyledonous and dicotyledonous plants. Analysis of ZmACR cis-acting elements revealed the optional involvement of ZmACR genes in light response, hormone response and stress resistance. Expression analysis of 8 ZmACR genes under salt treatment clearly revealed their role in the response to salt stress. Ectopic overexpression of ZmACR5 in Arabidopsis notably reduced salt tolerance compared to that of the wild type under salt treatment, suggesting that ZmACR5 has a negative role in the response to salt stress. Taken together, these findings confirmed the involvement of ZmACR genes in regulating salt stress and contributed significantly to our understanding of the molecular function of ACR genes in maize, facilitating further research in this field.

Genome-Wide Identification of GAST Family Members and Their Potential Roles in Epicotyl Dormancy in Chinese Cork Oak (Quercus variabilis).

Chinese cork oak (Quercus variabilis Blume) is a widespread tree species with high economic and ecological values. Chinese cork oak exhibits epicotyl dormancy, causing emergence heterogeneity and affecting the quality of seedling cultivation. Gibberellic acid-stimulated transcript (GAST) is a plant-specific protein family that plays a crucial regulatory role in plant growth, development, and seed germination. However, their evolution in Chinese cork oak and roles in epicotyl dormancy are still unclear. Here, a genome-wide identification of the GAST gene family was conducted in Chinese cork oak. Ten QvGAST genes were identified, and nine of them were expressed in seed. The physicochemical properties and promoter cis-acting elements of the selected Chinese cork oak GAST family genes indicated that the cis-acting elements in the GAST promoter are involved in plant development, hormone response, and stress response. Germinated seeds were subjected to gibberellins (GAs), abscisic acid (ABA), and fluridone treatments to show their response during epicotyl dormancy release. Significant changes in the expression of certain QvGAST genes were observed under different hormone treatments. QvGAST1, QvGAST2, QvGAST3, and QvGAST6 exhibited upregulation in response to gibberellin. QvGAST2 was markedly upregulated during the release of epicotyl dormancy in response to GA. These findings suggested that QvGAST2 might play an important role in epicotyl dormancy release. This study provides a basis for further analysis of the mechanisms underlying the alleviation of epicotyl dormancy in Chinese cork oak by QvGASTs genes.

Genome-wide characterization of MATE family members in common bean (Phaseolus vulgaris Linn.) and their expression profiles in response to abiotic stress

Multidrug and toxic compound extrusion (MATE) family plays an important role in the detoxification process of plants under various stresses. Nevertheless, a comprehensive study on MATE family members has thus far not been performed in common bean (Phaseolus vulgaris Linn.), limiting our understanding of its functions in common bean acclimation to stressful environments. In this study, a total of 56 PvMATEs members with uneven distribution on all chromosomes were identified from the common bean genome, with the most on chromosome 4 (10 members) and the least on chromosome 6 and 10 (1 members). Based on their orthologous relationship with those from Arabidopsis and rice, PvMATEs were classified into Clades I, II, III and Ⅳ, wherein 19, 15, 14, and 8 members were included, respectively. Gene ontology (GO) term and cis-regulatory element (CRE) analyses results indicated that the potential functions of PvMATEs were involved in hormonal response, developmental regulation and multiple stress response of common bean. The results of Phytozome and qRT-PCR analyze showed that the expression of PvMATEs in common bean are tissue specificity, with the most in root. Furthermore, 75 mM NaCl, 75 mM CuCl2, 0.35uM HgCl2, 16uM CdCl2, 4.38uM Benzo[c]-phenanthrene (C18H12) and 10 % PEG6000 treatments significantly up-regulated the expression levels of some PvMATEs in common bean roots, indicating that these genes are involved in multiple abiotic stresses respond of common bean. Taken together, these observations enriched our knowledge of MATE family in the common bean and benefit for breeding improved cultivars.

Down-regulating the stress axis: Living in the present while preparing for the future

The measurement of glucocorticoid (GC) hormones provides us with a window into the stress physiology of vertebrates and the adaptative responses they use to cope with predictable and unpredictable changes in the environment. Baseline GCs inform us about the metabolic demands they are subject to at that point in their yearly life-history stage, whereas GC changes (often increases) in response to acute challenges inform us on their capacity to cope with more immediate environmental challenges. However, baseline GC levels and the kinetics of GC responses to acute stressors can vary substantially among and within species, depending on individual characteristics (age, sex, condition, life-history stage). In addition, a thorough understanding of the stress status of an animal requires moving beyond the measurement of GCs alone by focusing on downstream measures of metabolic activation, such as oxidative stress. Here, we evaluated the changes in blood cortisol and oxidative stress markers in wild adult Columbian ground squirrels (Urocitellus columbianus), following a 30-min capture-handling stress performed in mid-late June. Measurements were taken when males were post-reproductive and preparing for hibernation and adult females were weaning litters. We found three key results. First, the time-course of GC increase was markedly slower (by an order of magnitude) than what is currently reported in the literature for most species of mammals, birds and reptiles. Second, there were marked differences in the male and female response, linked to differences in life-history stage: females close to weaning had abolished GC responses, whereas post-reproductive males did not. Third, there were mild to moderate increases in oxidative damage and decreases in oxidative defenses in response to our short-term challenge, consistent with the idea that short-term acute metabolic activation may carry physiological costs. However, these changes were not correlated to the changes in GCs, a novel result suggesting a disconnect between the hormonal stress response and oxidative damage.

Identification and expression analysis of the HSP70 gene family under abiotic stresses in Litchi chinensis

HSP70 protein, as an important member of the heat shock protein (HSP) family, plays an important role in plant growth, development, and response to biotic and abiotic stresses. In order to explore the role of HSP70 gene family members in Litchi chinensis under low temperature, high temperature, drought, and salt stress, bioinformatics methods were used to identify the HSP70 gene family members within the entire L. chinensis genome. The expression of these genes under various abiotic stresses was then detected using quantitative real-time PCR (qRT-PCR). The results showed that the LcHSP70 gene family consisted of 18 members, which were unevenly distributed across ten L. chinensis chromosomes. The LcHSP70 protein contained 479-851 amino acids, with isoelectric points ranging from 5.07 to 6.95, and molecular weights from 52.44 kDa to 94.07 kDa. The predicted subcellular localization showed that LcHSP70 protein was present in the nucleus, cytoplasm, endoplasmic reticulum, mitochondria, and chloroplast. Phylogenetic analysis divided the LcHSP70 proteins into five subgroups, namely Ⅰ, Ⅱ, Ⅲ, Ⅳ, and Ⅵ. The promoter regions of the LcHSP70 genes contained various cis-acting elements related to plant growth, development, hormone response, and stress response. Moreover, the expression of LcHSP70 genes displayed distint tissue-specific expression level, categorized into universal expression and specific expression. From the selected 6 LcHSP70 genes (i.e., LcHSP70-1, LcHSP70-5, LcHSP70-10, LcHSP70-14, LcHSP70-16, and LcHSP70-18), their relative expression levels were assessed under different abiotic stresses using qRT-PCR. The results indicated that the gene family members exhibited diverse responses to low temperature, high temperature, drought, and salt stress, with significant variations in their expression levels across different time periods. These results provide a foundation for further exploration of the function of the LcHSP70 gene family.

In silico analysis of the wheat BBX gene family and identification of candidate genes for seed dormancy and germination

BackgroundB-box (BBX) proteins are a type of zinc finger proteins containing one or two B-box domains. They play important roles in development and diverse stress responses of plants, yet their roles in wheat remain unclear.ResultsIn this study, 96 BBX genes were identified in the wheat genome and classified into five subfamilies. Subcellular localization prediction results showed that 68 TaBBXs were localized in the nucleus. Protein interaction prediction analysis indicated that interaction was one way that these proteins exerted their functions. Promoter analysis indicated that TaBBXs may play important roles in light signal, hormone, and stress responses. qRT-PCR analysis revealed that 14 TaBBXs were highly expressed in seeds compared with other tissues. These were probably involved in seed dormancy and germination, and their expression patterns were investigated during dormancy acquisition and release in the seeds of wheat varieties Jing 411 and Hongmangchun 21, showing significant differences in seed dormancy and germination phenotypes. Subcellular localization analysis confirmed that the three candidates TaBBX2-2 A, TaBBX4-2 A, and TaBBX11-2D were nuclear proteins. Transcriptional self-activation experiments further demonstrated that TaBBX4-2A was transcriptionally active, but TaBBX2-2A and TaBBX11-2D were not. Protein interaction analysis revealed that TaBBX2-2A, TaBBX4-2A, and TaBBX11-2D had no interaction with each other, while TaBBX2-2A and TaBBX11-2D interacted with each other, indicating that TaBBX4-2A may regulate seed dormancy and germination by transcriptional regulation, and TaBBX2-2A and TaBBX11-2D may regulate seed dormancy and germination by forming a homologous complex.ConclusionsIn this study, the wheat BBX gene family was identified and characterized at the genomic level by bioinformatics analysis. These observations provide a theoretical basis for future studies on the functions of BBXs in wheat and other species.

Identification and expression profiling of SAPK gene family members in Dendrobium officinale

To investigate the potential roles of stress-activated protein kinase (SAPK) gene family members in Dendrobium officinale, we employed multiple bioinformatics methods to identify the members of this family. The physicochemical properties, chromosomal localization, phylogenetic relationship, gene structure, and cis-acting elements of each D. officinale SAPK (DoSAPK) member were analyzed. In addition, their expression profiles in different tissues and under the low-temperature or salt stress treatment were determined by real-time fluorescence quantitative PCR. The results showed that D. officinale carried eight DoSAPK family members, which belonged to three groups (groups Ⅰ, Ⅱ, and Ⅲ). These genes were located on seven chromosomes, and there were two pairs of genes with replication. The DoSAPK members within the same group had similar gene structures, conserved motifs, and secondary structures. The cis-acting elements in the promoter regions of DoSAPK genes included abundant hormone and stress response elements. DoSAPK family members presented tissue-specific expression in D. officinale. Furthermore, they were differentially expressed under the low-temperature or salt stress treatment, which suggested that they might be involved in the responses to low-temperature and salt stress. Intriguingly, DoSAPK1 might play a role in the abiotic stress responses. The results laid a foundation for in-depth study of the members and roles of the DoSAPK gene family.