Programmable Cancer Subtype Evaluator via Multiply-Guaranteed Catalytic DNA Computing Circuit.

CDK4/6 inhibitors as neoadjuvant treatment in breast cancer—what can we learn?

Dual-miRNA-Propelled Three-Dimensional DNA Walker for Highly Specific and Rapid Discrimination of Breast Cancer Cell Subtypes in Clinical Tissue Samples

Fast and accurate identification of microbial pathogens is critical for the proper treatment of infections. Traditional culture‐based diagnosis in clinics is increasingly supplemented by metagenomic next‐generation‐sequencing (mNGS). Here, RNA/cDNA‐targeted sequencing (meta‐transcriptomics using NGS (mtNGS)) is established to reduce the host nucleotide percentage in clinic samples and by combining with Oxford Nanopore Technology (ONT) platforms (meta‐transcriptomics using third‐generation sequencing, mtTGS) to improve the sequencing time. It shows that mtNGS improves the ratio of microbial reads, facilitates bacterial identification using multiple‐strategies, and discovers fungi, viruses, and antibiotic resistance genes, and displaying agreement with clinical findings. Furthermore, longer reads in mtTGS lead to additional improvement in pathogen identification and also accelerate the clinical diagnosis. Additionally, primary tests utilizing direct‐RNA sequencing and targeted sequencing of ONT show that ONT displays important potential but must be further developed. This study presents the potential of RNA‐targeted pathogen identification in clinical samples, especially when combined with the newest developments in ONT.

To meet the growing demand for ultrasensitive diagnostics, representative hybrid platforms integrating nonenzymatic isothermal nucleic acid amplification such as catalytic hairpin assembly (CHA) with CRISPR/Cas systems have been developed. However, two major challenges remain: background leakage from spontaneous hairpin hybridization and inherent fluorescence from conventional ssDNA reporters. Here, we present a self-sustaining isothermal biosensing platform that addresses these limitations by combining CHA with a split activator-initiated CRISPR/Cas12a feedback circuit for the ultrasensitive detection of miRNA-155, a key biomarker of breast cancer. In our design, miRNA-155 initiates CHA to form a DNA duplex, which, along with the miRNA, acts as split activators to trigger CRISPR/Cas12a. Cas12a cleaves a ds-loop DNA reporter, releasing fluorescence and regenerating the target. This dual-recognition mechanism ensures strict target dependence, reduces background noise, and, with the reporter design, minimizes leakage. The released miRNA reactivates CHA, enabling continuous signal amplification through a self-sustaining feedback loop involving successive CHA and Cas12a trans-cleavage cycles, enhancing detection sensitivity. Via these features, the platform achieves attomolar sensitivity and excellent specificity, even distinguishing single-base miRNA variants. Direct detection of endogenous miRNA-155 in serum samples from breast cancer patients demonstrated clear differentiation from healthy controls. This strategy provides a robust molecular detection platform for the accurate and ultrasensitive detection of low-abundance miRNAs in biomedical studies.

Hand in hand catalytic hairpin assembly-based FÖrster resonance energy transfer biosensor for simultaneous detection of multiple MicroRNAs from breast cancer.

Highly sensitive and reliable detection of microRNA for clinically disease surveillance using SERS biosensor integrated with catalytic hairpin assembly amplification technology

An enzyme-free sensing platform for miRNA detection and in situ imaging in clinical samples based on DNAzyme cleavage-triggered catalytic hairpin assembly

First person profile: Dennis J. Slamon, MD, PhD: Dr. Slamon's pioneering work helped lead to the breakthrough breast cancer drug trastuzumab, which has saved and extended the lives of millions of patients.

Target-driven cascade amplified assembly of covalent organic frameworks on tetrahedral DNA nanostructure with multiplex recognition domains for ultrasensitive detection of microRNA

Ultrasensitive miRNA-21 detection via catalytic hairpin assembly-enhanced light-initiated chemiluminescence for early cancer diagnosis.

Purpose Pathological complete response (pCR) after neoadjuvant chemotherapy (NACT) for breast cancer (BC) is a prognostic factor for relapse-free and overall survival (OS). Despite recent therapeutic developments for early BC, there are still a significant proportion of patients who do not achieve pCR. In this study we established pCR rates in routine care and investigated possible predictive factors of pCR, including HER2 status. Methods We evaluated 980 stage I-III BC patients receiving NACT between 2013 and 2022. Patient characteristics, rates of pCR (ypT0-is ypN0), toxicities, treatment modifications and survival outcomes were recorded. Standard histopathological variables were recorded [with HER2-low status, defined as HER2 1+ on immunohistochemistry (IHC) or 2+ on IHC without in situ hybridization (ISH) gene amplification]. Overall survival (OS) and relapse-free survival (RFS) were calculated. Median follow-up time was 60.3 months (interquartile range 19.7-78.7). Results The mean age of the study population was 49.6 ± 11.2 years. 64% had stage II disease, 70.5% had grade 3 disease, and 90.5% had ductal histopathology. 34.6% had estrogen receptor (ER)-positive/HER2-negative, 27.1% had triple-negative (TN), and 38.3% had HER2-positive BC. 233 patients (23.8%) had HER2-low disease. pCR rate was 35.8% in the overall population, 16.8% in ER-positive/HER2 negative BC, 32.7% in TNBC and 55.2% in HER2-positive BC. pCR rate differed according to grade, histology, HER2 status, radiological and clinical response of disease (p < 0.001) and early discontinuation of NACT (p = 0.001), but not according to age, menopausal status, BRCA status, platinum use, NACT dose reduction or delays, neutrophils-to-lymphocytes, platelets-to-lymphocytes, or lymphocytes-to-monocytes ratios. OS and RFS were better following pCR [HR 0.23 (95% CI, 0.14-0.38, p < 0.001) and HR 0.27 (95% CI, 0.18-0.40, p < 0.001), respectively]. Among HER2-negative BC patients (605, 61.7% of the entire cohort), there was a trend to lower pCR rate in HER2-low compared to HER2-negative (IHC 0) BC (19.7% vs 26.3% respectively, p = 0.06). pCR rate was significantly lower in ER-positive/HER2-low compared to ER-positive/HER2-negative (IHC 0) BC (12.1% vs 20.9% respectively, p = 0.031), while no difference in pCR rates was observed in TNBC patients by HER2 status. Despite a lower rate of pCR, Kaplan-Meyer curve showed that OS was significantly better in the HER2-low BC compared to HER2-negative (IHC 0) population at 100-months follow up. Multivariable Cox-regression model in the HER2-negative BC cohort demonstrated that HER2-low status was an independent predictor of OS (HR 0.59, 95% CI, 0.39-0.89 p = 0.012). Conclusions In our real-world analysis, pCR rates are consistent with the published data. Many patients still have residual disease following NACT, predicting worse outcomes, and may benefit from further adjuvant systemic therapies. Consistent with other studies, our findings suggest a possible prognostic and predictive role of HER2-low status especially among patients with ER-positive BC. This could lay the foundations for novel therapies targeting HER2 among HER2-low cancer subtypes. Citation Format: Matilde Corianò, Nicolo M.L. Battisti, Hala Aziz, Nalinie Joharatnam Hogan, Antonino Musolino, Alistair Ring. Pathological complete response to neoadjuvant systemic therapy and its predictive factors among early breast cancer subtypes: the emerging role of HER2 [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 PO2-02-09.

Background AnnexinA2 (AnxA2) membrane deposition has a critical role in HB-EGF shedding as well as IL-6 secretion in breast cancer cells. This autocrine cycle has a major role in cancer cell proliferation, migration and metastasis. The objective of the study is to demonstrate annexinA2-mediated autocrine regulation via HB-EGF and IL-6 in Her-2 negative breast cancer progression. Methods Secretory annexinA2, HB-EGF and IL-6 were analysed in the peripheral blood sample of Her-2 negative ( n = 20) and positive breast cancer patients ( n = 16). Simultaneously, tissue expression was analysed by immunohistochemistry. The membrane deposition of these secretory ligands and their autocrine regulation was demonstrated using triple-negative breast cancer cell line model. Results Annexina2 and HB-EGF expression are inversely correlated with Her-2, whereas IL-6 expression is seen in both Her-2 negative and positive breast cancer cells. RNA interference studies and upregulation of annexinA2 proved that annexinA2 is the upstream of this autocrine pathway. Abundant soluble serum annexinA2 is secreted in Her-2 negative breast cancer (359.28 ± 63.73 ng/mL) compared with normal (286.10 ± 70.04 ng/mL, P < 0.01) and Her-2 positive cases (217.75 ± 60.59 ng/mL, P < 0.0001). In Her-2 negative cases, the HB-EGF concentrations (179.16 ± 118.81 pg/mL) were highly significant compared with normal (14.92 ± 17.33 pg/mL, P < 0.001). IL-6 concentrations were increased significantly in both the breast cancer phenotypes as compared with normal ( P < 0.001). Conclusion The specific expression pattern of annexinA2 and HB-EGF in triple-negative breast cancer tissues, increased secretion compared with normal cells, and their major role in the regulation of EGFR downstream signalling makes these molecules as a potential tissue and serum biomarker and an excellent therapeutic target in Her-2 negative breast cancer.

Hormone receptor-positive (HR+), HER2 negative (HER2−) breast cancer has a long-term risk of recurrence, compared with other breast cancer subtypes, which ultimately impacts mortality outcomes. Although adjuvant endocrine therapy plays an integral role in risk reduction, disparities persist—with Black women experiencing higher mortality rates compared to White women. Multiple studies have demonstrated that statin use can lead to a reduction in breast cancer mortality, but there is limited information in relation to the impact of statins among diverse patient populations in the United States. To assess whether statin use is associated with an improvement in breast cancer-specific mortality (BCSM), we conducted a large, observational study in women aged 66 and older diagnosed with Stage I–IIIC HR+, HER2- breast cancer between 2013 and 2017. We identified eligible women using the SEER-Medicare database and linked them to the BRIDGE (BReast cancer Investigation of Disparities in GEorgia) cohort, a survivorship cohort of the Georgia Cancer Registry. Patients needed to be continuously enrolled in Medicare Parts A, B, and D in the 12 months following a breast cancer diagnosis. We used Cox proportional hazards models to estimate the impact of statin use within 12 months of diagnosis on BCSM with adjustment for age, stage, and race using multivariable-adjusted hazard ratios (HRs). A total of 2483 participants were identified, and of those, 303 (12.2%) were statin users, 383 (15.4%) were non-Hispanic Black women, and 1564 (63%) resided in census tract with greater than or equal to 10% poverty levels at the time of diagnosis. Distribution of age, stage, and race were similar between statin and non-statin users. Of the statin users, 41/303 (13.5%) were non-Hispanic Black women. The majority of statin users were prescribed lipophilic statins alone (169; 55.8%), and atorvastatin was the most commonly prescribed statin. There was no association between statin use and BCSM (HR 1.09; 95% CI 0.62-1.90) in the multivariable-adjusted model. Although preliminary results suggest that there is no association between statin use and BCSM among women with Stage I–IIIC HR+, HER2− breast cancer, further analyses are planned to explore statin use as a time-varying exposure, which may provide additional insights into the potential relationship between statin use and BCSM. In addition, analyses examining potential heterogeneity by race, tumor and treatment characteristics, and comorbidities are forthcoming. Given the low cost and relative accessibility of statins, we anticipate these findings will inform the clinical management of women with operable breast cancer, increasing parity in outcomes and potentially narrowing race disparities. Citation Format: Ruth Sacks, Maya Bliss, Micah Streiff, Leah Moubadder, Maret Maliniak, Tsion Armidie, Lindsay Collin, Kevin Kalinsky, Lauren McCullough. The Impact of Statins on Breast Cancer Specific Mortality in a Diverse Patient Population with Stage I-III Hormone Receptor Positive, HER2 Negative 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 PO2-02-04.

An "on-off-on" self-powered biosensor based on enzyme-free amplification for detection of three biomarkers.

Alpha-foetoprotein (AFP) is taken as a diagnostic tumor marker for the screening and diagnosis of cancer. Nucleic acid-based isothermal amplification strategies are emerging as a potential technology in early screening and clinical diagnosis of AFP. The leakages between hairpins dramatically increase the background and reduce the sensitivity. Thus, it is necessary to develop some strategies to reduce the leakage for isothermal amplification strategies. A DNAzyme-locked leakless enzyme-free amplification system was developed for AFP detection in liver cancer and breast cancer. AFP could open the apt-hairpin and initiate the catalytic hairpin assembly (CHA) reaction to produce a Y-shaped duplex. Two tails of a Y-shaped duplex cleaved the two kinds of leakless hairpins. Then, the third tail of the Y-shaped duplex catalyzed the second CHA between the cleaved leakless hairpins to recover the fluorescent intensity. The limit of detection reached 5fg/mL by the two levels of signal amplifications. Importantly, the leakless hairpin design effectively reduced leakage between hairpins and weakened the background. In addition, it also showed a great promising potential for AFP detection in early screening and clinical diagnosis.