Negative Cells Research Articles

Efficacy of immune checkpoint inhibitors (ICIs) in PD-L1 negative Non-Small Cell Lung Cancer (NSCLC) - A meta-analysis based on reconstructed individual participant data.

Efficacy of immune checkpoint inhibitors (ICIs) in PD-L1 negative Non-Small Cell Lung Cancer (NSCLC) - A meta-analysis based on reconstructed individual participant data.

Elucidating the anticancer properties of posaconazole in triple-negative breast cancer through in-silico and in-vitro analysis.

Elucidating the anticancer properties of posaconazole in triple-negative breast cancer through in-silico and in-vitro analysis.

Green synthesis of gold nanoparticles from Dicoma anomala extract coated with liposome-encapsulated pheophorbide-a for enhanced photodynamic therapy against MDA-MB-231 triple negative breast cancer cells.

Green synthesis of gold nanoparticles from Dicoma anomala extract coated with liposome-encapsulated pheophorbide-a for enhanced photodynamic therapy against MDA-MB-231 triple negative breast cancer cells.

LINC01235 is an Upstream Regulator of the NFIB Gene and the NOTCH Pathway in Triple Negative Breast Cancer.

We identified a long non-coding RNA (lncRNA), LINC01235, with significant enrichment in luminal progenitor (LP)-like cells in triple negative breast cancer organoids and cell lines. Antisense-mediated knockdown or genetic knockout of LINC01235 in TNBC cell lines led to a decline in cell proliferation and adversely impacted the ability to form organoids. A comprehensive co-expression analysis, leveraging TCGA data, revealed a distinct correlation between LINC01235 expression and the expression of NFIB, a neighboring gene encoding a transcription factor. Subsequent CRISPR knockout or ASO-mediated knockdown studies demonstrated an upstream regulatory role of LINC01235 over NFIB. Moreover, our investigations demonstrated that LINC01235 regulates the NOTCH pathway through NFIB, and ChIRP-qPCR results indicated the direct binding of LINC01235 to the NFIB promoter. Our findings demonstrate that LINC01235 positively regulates NFIB transcription, which in turn modulates the NOTCH pathway, influencing LP-like cell proliferation in breast cancer progression. This study highlights a pivotal role of LINC01235 in TNBC and its potential as a therapeutic target. Implications: This study demonstrates the central role of LINC01235 as an upstream positive regulator of NFIB and the NOTCH signaling pathway to induce the production of luminal progenitor-like cells in TNBC.

The Vps13-like protein BLTP2 regulates phosphatidylethanolamine levels to maintain plasma membrane fluidity and breast cancer aggressiveness.

Lipid transport proteins (LTPs) facilitate non-vesicular lipid exchange between cellular compartments and have critical roles in lipid homeostasis. A recently identified family of bridge-like LTPs (BLTPs) is thought to form lipid-transporting conduits between organelles. One of these, BLTP2, is conserved across species but its function is not known. Here we show that BLTP2 regulates plasma membrane (PM) fluidity by increasing phosphatidylethanolamine (PE) levels in the PM. BLTP2 localizes to endoplasmic reticulum (ER)-PM contact sites, and transports PE in vivo, suggesting it drives PE movement from ER to PM. We find that BLTP2 works in parallel with another pathway that regulates intracellular PE distribution and PM fluidity. BLTP2 expression correlates with breast cancer aggressiveness. We found that BLTP2 facilitates growth of a triple negative breast cancer cell line and sustains its aggressiveness in an in vivo model of metastasis, suggesting maintenance of PM fluidity by BLTP2 may be critical for tumorigenesis in humans.

The effects of hsa-mir-26a-5p on cell proliferation, migration, and PI3K inhibitor sensitivity in metformin-resistant triple negative breast cancer cells

The effects of hsa-mir-26a-5p on cell proliferation, migration, and PI3K inhibitor sensitivity in metformin-resistant triple negative breast cancer cells

Evaluating follow-up in patients with lymph node-negative penile cancer at a high-volume centre.

To evaluate the current European Association of Urology/American Society of Clinical Oncology follow-up (FU) schedule with routine use of ultrasonography (US) ± US-guided fine-needle aspiration cytology (FNAC) in patients with lymph node (LN)-negative penile squamous cell carcinoma (PSCC). We evaluated FU outcomes for low-risk clinically LN-negative (cN0) and intermediate- to high-risk sentinel node (SN) negative (pathological N-stage [pN]0) patients with PSCC at a high-volume centre. We analysed routine inguinal US ± FNAC (in case of a suspicious LN) during FU. A competing risk analysis was performed to calculate the cumulative risk of LN metastases in these groups, with local recurrence as competing risk. A total of 201 patients with PSCC were analysed, with 2694 inguinal US investigations being evaluated. FNAC was performed during 270 US procedures (10.0%). A LN metastasis was found in four of 270 US + FNAC procedures (1.5%, 0.2% of all US). All tumours were intermediate- to high-risk tumours. Three occurred within the first year after primary treatment, and one at 20 months. The cumulative risk of developing LN metastases in cN0 low-risk tumours was 0% (no events occurred) and in SN-negative intermediate- to high-risk group 2.0%, both at 1 year of FU. Two (out of four) patients with LN recurrence died from disease. After adequate staging and treatment, patients with cN0/pN0 PSCC rarely develop LN metastasis during FU. As these metastases generally occur within 1 year after treatment, we recommend (3-monthly) routine inguinal US only during the first year of FU. Thereafter, the frequency of FU visits and examinations can be reduced in patients capable of self-examination of the penis, resulting in a substantial (up to a 50%) reduction of FU visits compared to current guideline recommendations.

A novel bispecific integrin α5β1/αv antibody reprograms the Myc-regulated basal phenotype of prostate cancer with natural killer cell-mediated tumor elimination.

Integrin α5β1 and αv crosstalk in chemotaxis and clonogenic survival of prostate cancer cells is abrogated by a bispecific α5β1/αv antibody (BsAbα5β1/αv), which uniquely induces internalization and lysosomal degradation of target integrins. We hypothesized that the BsAbα5β1/αv inactivates pathological mechanosignaling pathways that correlate with integrin expression from patient samples. Mechanistic studies indicate that the BsAbα5β1/αv uniquely reverses YAP, beta-catenin and FAK nuclear localization compared to monospecific integrin α5β1 and αv antibody controls in basal-type androgen-receptor negative prostate cancer cells. Dual integrin αv and α5 knockdown alone phenocopied the BsAbα5β1/αv effect. Following BsAbα5β1/αv treatment, ATAC-seq studies indicated the chromatin accessibility to TEAD and AP-1 family members was markedly reduced. In vitro and in vivo RNA-seq indicated down-regulation of Myc/E2F, TGF-beta and epithelial mesenchymal transition (EMT) and upregulation of Type I and II interferon transcriptomic pathways. The BsAbα5β1/αv induced CXCL10 and CCL5 cytokine secretion, immune-infiltration of tumors, and natural-killer cell-mediated elimination of the basal-type prostate cancer xenografts in nude mice. αv integrin was highly expressed and principally correlated with the Myc signaling pathway in rapid autopsy tissue microarrays, consistent with correlative data from the SU2C metastatic castration-resistant prostate cancer and DKFZ early-onset prostate cancer cohorts. These studies connect integrin signaling with the central biology of basal-type and castration-resistant prostate cancer and define a novel therapeutic strategy that controls critical immunosuppressive pathways. Implications: Dual integrin α5β1/αv targeting with a bispecific antibody represents a novel therapeutic strategy that reprograms the epigenetic and transcriptomic signature of basal-type prostate cancer with induction of immunological tumor control.

Yttrium oxide nanoparticles induce selective cytotoxicity, genomic instability and ROS mitochondrial P53 mediated apoptosis in human pancreatic cancer cells

Pancreatic cancer is a hard-to-treat tumor with a poor prognosis. While traditional pancreatic cancer therapies can be effective, issues like cytotoxicity, low selectivity, and drug resistance still pose major challenges. Nanotechnology has shown promise in improving cancer diagnosis and treatment. Yttrium oxide nanoparticles (Y2O3-NPs), for example, have demonstrated potent selective cytotoxicity against triple negative breast cancer cells; but their effects on pancreatic cancer cells have not been explored. This study aimed to explore the impact of Y2O3-NPs on cell proliferation, DNA integrity, and oxidative stress in pancreatic cancer (PANC-1) and human skin fibroblast (HSF) cells. The cytotoxicity of Y2O3-NPs after 72 h were estimated using Sulforhodamine (SRB) cytotoxicity assay, while alkaline Comet assay was done to study genomic DNA integrity. Generation level of reactive oxygen species (ROS) and integrity of mitochondrial membrane potential were also analyzed. Apoptosis induction was investigated using Flow Cytometry and expression level of apoptotic (p53), anti-apoptotic (Bcl2) and mitochondrial (ND3) genes was measured using quantitative RTPCR. Our findings exhibited that Y2O3-NPs had strong selective cytotoxicity against PANC-1 cells with an IC50 value of 31.06 µg/ml, while having minimal effect on normal HSF cells (IC50 = 319.21 µg/ml). Treatment of PANC-1 cells with Y2O3-NPs at the IC50 concentration for 72 h significantly increased intracellular ROS levels and DNA damage, along with a notable reduction in mitochondrial membrane potential. Additionally, a significant rise in necrotic, early, and late apoptotic cells was observed, accompanied by downregulation of the anti-apoptotic Bcl2 gene and upregulation of the apoptotic p53 and mitochondrial ND3 genes. These findings highlight the selective toxicity of Y2O3-NPs towards cancerous PANC-1 cells, with minimal impact on normal cells. Y2O3-NPs appear to induce apoptosis in cancer cells by increasing ROS generation, damaging DNA, disrupting mitochondrial function, and triggering cell death. This study suggests that Y2O3-NPs may be a promising candidate for pancreatic cancer treatment. Further research is needed to fully explore their therapeutic potential.

Antimigratory and Antiproliferative Effects of the Brain-Targeted Peptide Conjugate PepH3-vCPP2319 on Triple Negative Breast Cancer Cell Cultures.

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype affecting mostly younger women with a poor 5-year overall survival. It is characterized by a high metastization rate, particularly to the brain, where the blood-brain barrier (BBB) hinders the pharmaceuticals delivery. New anticancer drugs able to inhibit cell migration are required to effectively prevent the development of metastasis. PepH3-vCPP2319 (AGILKRW(Ahx)WRRRYRRWRRRRRQRRRPRR-amide), consisting of the conjugation of the BBB peptide shuttle (BBBpS) PepH3 (AGILKRW-amide) to the anticancer peptide (ACP) vCPP2319 (WRRRYRRWRRRRRQRRRPRR-amide), was reported to have high anticancer activity (IC50 = 5.0 μM) toward highly aggressive TNBC cells (MDA-MB-231) paired with 2-fold increased accumulation in the brain when compared to unconjugated vCPP2319. Herein, we demonstrate that PepH3-vCPP2319 inhibits cell migration and proliferation in wound healing assays, outperforming the gold standard small chemical inhibitor, iCRT-3. The concentration required to inhibit cell migration is 10-fold lower for PepH3-vCPP2319 (0.5 μM) when compared with iCRT-3 (50 μM). Likewise, PepH3-vCPP2319 at 2.5 μM was more efficient in preventing cell proliferation when compared with 50 μM iCRT-3, with 45% reduction in spheroid diameter. This study sheds light on the antimetastatic potential of PepH3-vCPP2319 through abrogation of cell migration to distant sites, including the brain.

ASO Visual Abstract: Thoracic Adipose Volume is Associated with Occult Nodal Disease- and Disease-Free Survival in Females with Clinically Node Negative Non-Small Cell Lung Cancer.

ASO Visual Abstract: Thoracic Adipose Volume is Associated with Occult Nodal Disease- and Disease-Free Survival in Females with Clinically Node Negative Non-Small Cell Lung Cancer.

Abstract P4-05-27: Comparative analysis of TROP2 expression in tumor tissues and circulating tumor cells (CTCs) in the peripheral blood of patients with triple negative breast cancer

Abstract Background: Trophoblast cell surface antigen 2 (TROP2) promotes breast cancer (BC) development, invasion and metastasis, with promising role as a biomarker and therapeutic target in triple negative BC. Due to the dynamic tumor evolution during disease progression, a significant discordance in tumor cell profiles is frequently observed among primary tissues and distant metastases. In this regard, analyses of circulating tumor cells (CTCs) in the peripheral blood (PB) can inform on the expression of biomarkers in real-time. In the current study we assessed in parallel the expression of TROP2 on CTCs and matched primary tumors and metastatic sites from patients with triple negative BC. Methodology: PB was collected from 54 patients and CTCs were enriched by ficoll-density gradient centrifugation. Cytospins were immunofluorescently stained using antibodies for cytokeratins (Clones: AE1/AE3 & C11), CD45 and TROP-2 (Enzo Life Sciences); TROP2 expression on CTCs was defined as high, low or negative, by using the high TROP2-expressing MDA.MB.231 triple negative BC cell line as internal control. Matched primary (n=51) and metastatic (n=7) tumor tissue samples were evaluated for TROP-2 expression by immunocytochemistry (IHC); H-score was calculated as follows: (3 × % cells with strong intensity staining) + (2 × % cells with moderate intensity staining) + (1 × % cells with mild intensity staining), ranging from 0 to 300, and the following expression categories were defined: H-score 0 to <100: TROP2 low; H-score 100-200: TROP2 medium; H-score >200-300: TROP2 high. Results: CTCs (CK+/CD45- cells) were identified in 12 out of 54 patients evaluated (total CTC counts: n=80; mean CTC counts per patient: n=6.7). TROP2-expressing CTCs were detected in 75% of CTC-positive patients and represented the 95% of total CTCs. Specifically, high and low TROP2-expressing CTCs were identified in 66.7% and 41.7% of patients, representing the 81.3% and 13.7% of total CTCs, respectively. Differential TROP2 expression levels (high, medium and low) were also observed in both primary and metastatic tumors, showing a great intra-tumoral heterogeneity. High TROP2 expression was identified in 58.8% and 57.1% of primary and metastatic tissues, respectively. When matched primary and metastatic tissues were analyzed, a decrease in TROP2 expression was observed [median H-Score: 172.5 (range: 11-300) versus 87.5 (range: 5-150) in primary and metastatic tissue, respectively, p=0.068)]. CTC detection in the PB was not associated with TROP2 expression levels in primary or metastatic tissue (CTCs were identified in 13.3% and 50% of patients with high TROP2-expressing primary and metastatic tumors, respectively). Finally, there was no concordance in TROP2 expression pattern among CTCs and the respective tumor (high, low and negative TROP2-expressing CTCs were identified in 50%, 25% and 25% of patients with high TROP2-expressing primary tumors, respectively; high and low TROP2 -expressing CTCs were evident in all patients with high TROP2-expressing metastatic tumors). Conclusions: Herein we demonstrate for the first time a significant discrepancy in TROP2 expression among CTCs, primary and metastatic tumor tissue samples in triple negative BC. A lower TROP2 expression was observed in metastatic as compared to primary tissue, while no concordance was demonstrated among CTCs and the respective tumors. The results suggest the dynamic change in TROP2 expression status among different disease sites, thus highlighting the value of using liquid biopsy as a tool for real-time biomarker assessment in triple negative BC. Citation Format: Dimitrios Mavroudis, Eleni Lagoudaki, Sofia Gounaki, Sofia Hatziavraam, Charalampos Fotsitzoudis, Kleita Michaelidou, Sofia Agelaki, Maria A Papadaki. Comparative analysis of TROP2 expression in tumor tissues and circulating tumor cells (CTCs) in the peripheral blood of patients with triple negative breast cancer [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr P4-05-27.

Abstract P1-07-26: Annexin A6 modulates the secretion of pro-inflammatory cytokines and exosomes via membrane fusion with SNAP 23 in triple negative breast cancer

Abstract The role of Annexin A6 (AnxA6) in the secretion of pro-inflammatory cytokines (PICs) and exosomes in triple negative breast cancer cells remains to be fully elucidated. However, understanding how AnxA6 influences or whether it is involved in the secretion of PICs and exosomes is still debatable. PICs are largely known to be secreted into the cancer microenvironment through exocytosis. Moreover, previous studies have reported the involvement of PICs in drug resistance and metastasis during exocytosis. We had previously shown that AnxA6 is secreted as a component of extracellular vesicles and that breast cancer cells secrete annexins via the exosomal pathway. Here, we examined the secretory function of AnxA6 in monocyte chemoattractant protein 1 (MCP-1) and interleukin 8 (IL-8) cytokines that contribute to triple negative breast cancer (TNBC) progression and we show that the secretion of cytokines and exosomes is AnxA6-dependent. AnxA6 upregulation in TNBC cells promotes increased extracellular vesicles (EVs) secretion. We equally show that cholesterol loading into EVs may also be AnxA6-dependent and that lapatinib-induced upregulation of AnxA6 leads to increased cytokine secretion in MDA-MB-468 lapatinib-resistant (LAP-R) cells. Furthermore, we demonstrate that lapatinib-induced upregulation of AnxA6 expression in EVs in MDA-468 LAP-R cells also leads to increased cholesterol accumulation. SNAP 23 interacts with AnxA6 for increased secretion through fusion of vesicles with plasma membrane. Together, these data not only suggest that PIC, exosome secretion, and cholesterol enrichment is AnxA6-dependent, but also provide evidence that induced AnxA6 upregulation triggered by chronic lapatinib treatment will lead to increased PIC secretion in the cells, which is bad for TNBC progression and poor patient outcome. Citation Format: Nobelle Sakwe, Portia Thomas, Josiah Ochieng, Amos M. Sakwe. Annexin A6 modulates the secretion of pro-inflammatory cytokines and exosomes via membrane fusion with SNAP 23 in triple negative breast cancer [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr P1-07-26.

Abstract P4-10-26: PRISM high-throughput screening to elucidate how tumor microenvironments modulate drug responses of breast cancer cells

Abstract Cancer treatment options including small molecules, antibody-drug conjugates (ADCs), CAR T-cells, and combination therapies are rapidly expanding to improve cancer patient survival. However, efficacy of targeted therapies is limited by interpatient and intratumoral heterogeneity in breast cancer, and may be further exacerbated by dysregulated tumor microenvironments. For example, studies suggested that changes in tumor and breast tissue microenvironment in obesity (e.g. inflammation, increased growth factors) could lead to worsened prognosis that could be related to therapy resistance. Systematic characterization of the cellular effects of emerging drug candidates, particularly across different tumor microenvironments, is challenging, as it needs to be surveyed in diverse cancer contexts beyond just a handful of cancer cell lines. To overcome this challenge, we created PRISM (Profiling Relative Inhibition Simultaneously in Mixtures), an effective drug discovery screening platform with over 900 genetically distinct cancer cell lines of diverse lineages, to rapidly test therapeutics in a high-throughput manner. These cell lines, including 33 different breast cancer cell lines covering diverse cell types and subgroups, carry a specific 24 nucleotide “barcode” tag and have rich baseline genomic and functional characterization. They are first pooled into 20-25 cell pools of different lineages with similar growth rates, then subsequently cultured together with or without drugs. The relative barcode quantity read out by NextGen sequencing determines cell line responses to drugs. This pooled approach significantly reduces the time and the resources required for drug screening. Importantly, utilizing the rich cell line characterization to interpret viability profiles enables the identification of drivers of differential sensitivity and potential biomarkers of compound response. Furthermore, we created a breast cancer cell specific pool to investigate how media conditions mirroring different tumor microenvironments [e.g. media filtration, addition of insulin (0.01 mg/mL) or EGF (0.01 ug/mL)] can impact drug effects on breast cancer cells. In the PRISM 900+ cell panel treated with ADCs conjugated to different payloads that target HER2, there was a clear association between cell line sensitivity and ERBB2 (encoding HER2) gene expression, with the strongest effects on viability observed in a subset of cell lines overexpressing ERBB2. Upon further comparison of biomarker profiles across different ADCs, we established relationships between payload resistance and ABCB1 efflux pumps emerged for a subset of ADC payloads. Using the breast cancer cell specific pool, our data showed that estrogen receptor (ER) degraders fulvestrant and vepdegestrant (ARV-471) had significantly greater effects in reducing viability of ER+ versus ER- breast cancer cell lines. Media filtering reduced the drug effects of fulvestrant but not ARV-471, suggesting that ARV-471 efficacy may be less sensitive to the tumor microenvironment than fulvestrant. Lapatinib (HER2/neu and EGF receptor inhibitor) was more effective in killing HER2+ than HER2- negative breast cancer cells, with cells cultured in EGF-spiked media being more sensitive to lapatinib, whereas media filtration diminished its effects. We did not observe breast cancer subgroup specific killing by alpelisib (PI3K inhibitor) or olaparib (PARP inhibitor), but breast cancer cells became less sensitive to alpelisib when cultured in either insulin- or EGF-spiked media, and less sensitive to olaparib in EGF-spiked media. Overall, our study highlights how the PRISM 900+ cell panel can provide insights into drug specificity, cancer subtype selectivity, and to uncover clinically relevant targets. The breast cancer specific pool can further increase the throughput of understanding how drug responses may be altered in different tumor microenvironments in breast cancer. Citation Format: Blanche C. Ip, Jillian N. Eskra, Tenzin Sangpo, Ashish Bino George, Melissa Ronan, Matthew G. Rees, Jennifer A. Roth. PRISM high-throughput screening to elucidate how tumor microenvironments modulate drug responses of breast cancer cells [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr P4-10-26.

Dose Optimization of ClpP Agonists Using an In Vitro Microfluidic Perfusion Platform and In Silico Pharmacokinetic-Pharmacodynamic Modeling.

Small molecule activators of the mitochondrial caseinolytic protease P (ClpP agonists) can disrupt tumor metabolism and deprive tumors of their energy needs. The imipridone, ONC201, is a ClpP agonist currently undergoing clinical evaluation across multiple cancer types, while additional analogs with improved potency and selectivity are in preclinical development. Preclinical studies in mice have demonstrated a unique pharmacokinetic-pharmacodynamic (PK-PD) relationship for ONC201 characterized by prolonged pharmacology following a single dose. This motivated the selection of an initial human dosing regimen of every three weeks, and subsequent dose exploration studies in mice led to dose intensification in human patients. However, a systematic analysis of ClpP agonist PK-PD relationships has not been performed, and the optimal exposure profile for ClpP agonists remains undefined. To address this gap, we combined PK-PD modeling with a microfluidic perfusion platform as an animal-alternative approach for translational PK-PD of ClpP agonists. We demonstrate that the anti-proliferative effect on triple negative breast cancer cells correlates with the magnitude and duration of ClpP agonist exposure above a threshold concentration required for ClpP activation. Moreover, we demonstrate that PK-PD model simulations using parameters derived from microfluidic perfusion datasets can successfully predict the anti-tumor efficacy of a ClpP agonist in a mouse tumor xenograft study. These studies support the translational relevance of the animal-alternative in vitro PK-PD platform and its utility to help guide dose optimization of ClpP agonists as cancer therapeutics.

Optimal design methodology for negative stiffness structures considering lifespan and additive manufacturing uncertainty

Abstract Negative stiffness honeycomb unit cells (NSH-UCs), which employ negative stiffness beams (NSBs), are capable of absorbing impact energy and are reusable, making them promising for energy-absorbing structural applications. However, limited research has addressed their operational lifespan and manufacturing variability, both of which are critical for practical implementation. This study aims to establish a design optimization framework for NSH-UCs that accounts for both performance metrics—such as energy absorption—and operational aspects like reusability and manufacturing-induced uncertainty. To this end, NSH-UC samples with varying dimensions were fabricated using fused filament fabrication (FFF) with PLA/PHA filaments, and their mechanical behavior was evaluated through quasi-static and cyclic compression tests. A surrogate-based optimization method was then applied to improve energy absorption and extend service life, while considering geometric and material uncertainties inherent to the additive manufacturing process. The proposed framework led to a significant improvement in energy absorption (EA) and end-of-life (EOL) performance compared to the initial design, despite only modest changes in specific energy absorption (SEA). These findings demonstrate the feasibility of incorporating performance, reliability, and manufacturing variability into early-stage design, highlighting the framework’s potential for structural health monitoring (SHM) and prognostic health management (PHM) applications.

In Vitro Effect of Ferruginol, Tanshinone, and Carnosol Analogues on the Proliferation of Three Breast Cancer Cell Lines

Ferruginol, tanshinones and carnosol are considered privileged natural products due to their demonstrated diverse biological activities with relevance to cancer research. Globally, cancer continues to be a major contributor to mortality rates, making these compounds potentially valuable molecular scaffolds for further development as potential anticancer agents. In this work, a focused library of ferruginol, tanshinone IIA, and carnosol analogues was studied to examine their effectiveness against various solid tumor models. The compounds were efficiently synthesized from either methyl 12-hydroxy-dehydroabietate or 12-hydroxydehydroabietylamine in 1–3 step processes with good chemical yields. The compounds that were synthesized underwent a methodical evaluation using multiple biological tests (including viability assays, clonogenic assays, and mitochondrial membrane polarization measurements) to determine their ability to inhibit in vitro the growth of three breast cancer cell linages. It was determined that while most compounds exhibited biological activity, compounds 10 and 11 demonstrated significant efficacy against triple negative breast cancer cells. These compounds continue to show promising biological activity, suggesting that additional studies to understand their mechanisms of action would be valuable.

Dual inhibition of EGFR and PI3Kinase signaling in EGFR amplified triple negative breast cancer cells induces apoptosis and reduces tumor growth

Background:Epidermal Growth Factor Receptor (EGFR) family signaling is commonly dysregulated in cancer by amplification or activating mutations. We investigated the incidence of EGFR amplification with or without PI3Kinase pathway mutations in breast cancer and whether patients with genetic abnormalities in both pathways can be targeted by dual EGFR/PI3K inhibition.Methods:EGFR amplification and PI3K pathway mutations were studied through data sets from cBioPortal and Caris. We used the triple negative breast cancer (TNBC) cell lines BT20 (amplified EGFR, PIK3CA activating mutation), MDA-MB-468 (amplified EGFR, PTEN deletion), and MDA-MB-231 (no amplification of EGFR or PI3K pathway mutations). EGFR amplification was determined by immunoblot and fluorescent in situ hybridization, and PI3K mutations by sequencing. Signaling was determined by immunoblot, drug synergy by cell viability, cell death by propidium iodide staining, cell cycle analysis by flow cytometry, and animal studies through xenografts.Results:EGFR amplification is found in approximately 1–2.5% of breast cancer patients, more frequently in TNBC (2.45–6.7%) and ER-/HER2+ (1.3–6.5%) breast cancers, and in the molecular basal (2.33–8.1%) and HER2 enriched (1.87–5.4%) subtypes. Overall survival is shorter among patients compared to those with unamplified EGFR. Up to 71% of EGFR amplified tumors have activating mutations in the PI3K pathway. Combination of EGFR and PI3K inhibitors more dramatically reduced mTOR and AKT signaling in the BT20 and MDA-MB-468 cells, whereas the inhibition of downstream signaling was less significant in MDA-MB-231 cells. Combination of EGFR and PI3K inhibitors reduced cell viability in these three cell lines, but inhibition was greater, statistically significant, and synergistic in MDA-MB-468 and BT20 compared to MDA-MB-231. Only MDA-MB-468 and BT20 cells had an increased fraction of apoptotic cells. EGFR or PI3K inhibition alone in a BT20 xenograft model reduced tumor volume, however the combination was the only statistically significant reduction in tumor volume when compared to vehicle control.Conclusions:EGFR/PI3K inhibitor combination causes apoptosis and reduction in tumor growth in cells with EGFR amplification and PI3K alteration. Dual inhibition of EGFR/PI3K presents as a potential targeted therapy in patients with EGFR amplification and aberrant PI3K signaling.

Isolation and characterization of human cKIT positive amniotic fluid stem cells obtained from pregnancies with spina bifida

Myelomeningocele (MMC) and myeloschisis (MS) are considered as the most severe forms of spina bifida aperta characterized by incomplete closure of the neural tube during the first trimester of pregnancy. Open maternal-fetal surgery has been identified as a promising alternative for the repair of MMC/MS. The main goal of this study was the isolation and characterization of stem cells derived from MMC/MS amniotic fluid samples. Human amniotic fluid samples were obtained from pregnant women who underwent surgery for fetal spina bifida repair. We applied fluorescence activated-cell sorting (FACS) to immunoselect stem cells from heterogeneous populations of amniocytes based on cKIT (CD117) expression. The cKIT+ amniocytes were then contrasted with cKIT− and unselected amniocytes in order to characterize the phenotype of these cells. cKIT-expressing amniocytes exhibited spindle-shape morphology, while amniocytes negative for cKIT were round-shaped. qRT-PCR analysis revealed significant upregulation of mean mRNA levels of KIT and CD90 genes and downregulation of CK8 gene in cKIT-expressing amniocytes, compared to the negative and unsorted cells. The expression of pluripotency antigens was comparable in sorted cKIT+ and cKIT− amniocytes and in unselected cells. Our results are of pivotal importance for the future application of amniotic fluid derived stem cells in spina bidfida repair.

Reshaping the landscape of locoregional treatments for breast cancer liver metastases: A novel, intratumoral, p21-targeted percutaneous therapy increases survival in BALB/c mice inoculated with 4T1 triple negative breast cancer cells in the liver

Patients with disseminated metastatic disease from breast cancer are likely to have liver involvement in >50% of cases at some point during disease progression. These patients have a poor prognosis; and, when treated with the standard of care systemic therapy they have a median survival of <9-months. Increasing survival in breast cancer patients will likely require the administration of better therapies that are specifically targeted to treat distant metastases. One approach to increasing treatment efficacy for breast cancer liver metastases is through the application locoregional therapies. Locoregional therapies are an appealing interventional approach for breast cancer patients with liver metastases since these tumor lesions are accessible via minimally invasive procedures that can be administered using either ultrasound or CT imaging. Current locoregional therapies to treat breast cancer liver metastases are non-specific and have not produced significant increases in survival. The goal of this study was to design and test a targeted locoregional therapeutic intervention for breast cancer liver metastases. The lead candidate, a fixed-dose small-molecule drug called MBC-005, was tested in vitro and then the efficacy was evaluated in a BALB/c mouse liver metastases model. A novel formulation of N-allyl noroxymorphone hydrochloride incorporated into an alginate-based gel overcomes many of the limitations associated with the administration of small-molecule drugs, which include solubility, off-target toxicity, and enzymatic degradation. In vitro results demonstrated that MBC-005 mediated its anti-tumorigenic effect through a p21-dependent mechanism via a novel molecular pathway, in which N-allyl noroxymorphone component of MBC-005 stimulated the opioid growth factor receptor to increase p21 expression. Intratumoral administration of MBC-005 increased survival 3.9-fold in mice and significantly decreased tumor volume 4-fold. While many cytotoxic therapies increase p21 expression as a response to DNA damage, MBC-005 increased p21 expression independent cytotoxic DNA damage. MBC-005 did not induce off-target toxicity; and, as such, would be amenable to multiple rounds of administration. Nevertheless, it is notable that the positive effects of MBC-005 treatment on increasing survival and decreasing tumor volume in mice was achieved using a single dose.