Expression Of ErbB Receptors Research Articles

Breast cancer patient-derived organoids for the investigation of patient-specific tumour evolution

BackgroundA reliable preclinical model of patient-derived organoids (PDOs) was developed in a case study of a 69-year-old woman diagnosed with breast cancer (BC) to investigate the tumour evolution before and after neoadjuvant chemotherapy and surgery. The results were achieved due to the development of PDOs from tissues collected before (O-PRE) and after (O-POST) treatment.MethodsPDO cultures were characterized by histology, immunohistochemistry (IHC), transmission electron microscopy (TEM), scanning electron microscopy (SEM), confocal microscopy, flow cytometry, real-time PCR, bulk RNA-seq, single-cell RNA sequencing (scRNA-seq) and drug screening.ResultsBoth PDO cultures recapitulated the histological and molecular profiles of the original tissues, and they showed typical mammary gland organization, confirming their reliability as a personalized in vitro model. Compared with O-PRE, O-POST had a greater proliferation rate with a significant increase in the Ki67 proliferation index. Moreover O-POST exhibited a more stem-like and aggressive phenotype, with increases in the CD24low/CD44low and EPCAMlow/CD49fhigh cell populations characterized by increased tumour initiation potential and multipotency and metastatic potential in invasive lobular carcinoma. Analysis of ErbB receptor expression indicated a decrease in HER-2 expression coupled with an increase in EGFR expression in O-POST. In this context, deregulation of the PI3K/Akt signalling pathway was assessed by transcriptomic analysis, confirming the altered transcriptional profile. Finally, transcriptomic single-cell analysis identified 11 cell type clusters, highlighting the selection of the luminal component and the decrease in the number of Epithelial–mesenchymal transition cell types in O-POST.ConclusionNeoadjuvant treatment contributed to the enrichment of cell populations with luminal phenotypes that were more resistant to chemotherapy in O-POST. PDOs represent an excellent 3D cell model for assessing disease evolution.

Abstract 6899: Enfortumab vedotin (EV): Efficacy comparison with trastuzumab deruxtecan (T-DXd) and sacituzumab govitecan (SG) and its potential for combination benefit in bladder cancer XPDX models

Abstract EV is a Nectin-4 targeting antibody-drug conjugate (ADC) with an MMAE payload, recently approved for bladder cancer treatment. We established and characterized 18 bladder XPDX models representing primary and metastatic disease from naïve or clinically treated patients, some with actionable mutations. To better understand the potential additive benefit of EV in bladder cancer, we evaluated the ADC alone and in combination with agents targeting ERBB2, PIK3CA or FGFR3. In addition, we tested EV alone in ST5420B, a model established from a patient who progressed on EV after five cycles, as well as ST975B, a model which harbors a novel NECTIN4 fusion. 18 bladder XPDX models were established in immune-deficient mice, including 5 from newly diagnosed patients (2 from primary site), and 13 from recurrent disease (2 from primary site). Nectin-4, Trop2 protein and ERBB2 receptor expression was tested, and models profiled using WES and RNAseq. For in vivo studies, all models were evaluated against cisplatin, T-DXd, SG, and EV. Models with actionable mutations were tested with alpelisib, erdafitinib, and neratinib alone and in combination with EV. Endpoints in all studies included tumor volume and time from treatment initiation with %T/C values and tumor regression reported at study completion; a %T/C of ≤ 20% versus control was considered sensitive. Tumor regression (%T/C<0%) versus Day 0 tumor volume was also reported. Most models stained positive for Nectin-4 and Trop-2. FGFR3 variants were found in five models and ERBB2 mutations in two while several models reported PIK3CA variants; deletions in RB1 and CDKN2A were common. In vivo studies reported differential responses to cisplatin with the most sensitive models established from treatment-naïve patients. EV and SG tested alone reported activity in several models; EV added benefit in some combination treatments. We have characterized a panel of bladder XPDX models and benchmarked them against T-DXd, EV and SG alone and EV in combination with targeted therapies. This data is a valuable tool in further developing EV and identifying novel therapies for bladder cancer. Citation Format: Johnnie Flores, Heaven Sessions, Alyssa Simonson, Natalia Banos, Tahmineh Rouzbahani, Armando Diaz III, Jim Lund, Emiliano Calvo, Victor Moreno, Kyriakos Papadopoulos, Drew Rasco, Amita Patnaik, Scott Ulmer, Luis Rodriguez, Michael Wick. Enfortumab vedotin (EV): Efficacy comparison with trastuzumab deruxtecan (T-DXd) and sacituzumab govitecan (SG) and its potential for combination benefit in bladder cancer XPDX models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6899.

The ErbB Signaling Network and Its Potential Role in Endometrial Cancer.

Endometrial cancer (EC) is the second most common malignancy of the female reproductive system worldwide. The updated EC classification emphasizes the significant role of various signaling pathways such as PIK3CA-PIK3R1-PTEN and RTK/RAS/β-catenin in EC pathogenesis. Some of these pathways are part of the EGF system signaling network, which becomes hyperactivated by various mechanisms and participates in cancer pathogenesis. In EC, the expression of ErbB receptors is significantly different, compared with the premenopausal and postmenopausal endometrium, mainly because of the increased transcriptional activity of ErbB encoding genes in EC cells. Moreover, there are some differences in ErbB-2 receptor profile among EC subgroups that could be explained by the alterations in pathophysiology and clinical behavior of various EC histologic subtypes. The fact that ErbB-2 receptor expression is more common in aggressive EC histologic subtypes (papillary serous and clear cell) could indicate a future role of ErbB-targeted therapies in well-defined EC subgroups with overexpression of ErbB receptors.

Clinicopathological significance of concurrent ErbB receptor expression in human meningioma.

In general, human meningiomas grow slowly and have a favourable prognosis; however, some are prone to recur despite their benign histology. Therefore, knowledge of their tumour biology is essential to determine objective biomarkers that can identify cases with an increased risk for recurrence and to generate effective treatment options. Thus, studies on the epidermal growth factor receptor (EGFR) family, comprising ErbB1/EGFR, ErbB2/HER2, ErbB3/HER3 and ErbB4/HER4, are important. We have recently published papers on the expression of each of these receptor proteins in human meningiomas. The present study aimed to assess the clinicopathological significance of their concurrent expression. A total of 185 grade 1 and 2 meningiomas with robust clinical data underwent immunohistochemical analyses with antibodies against the aforementioned receptors. All meningiomas exhibited upregulation of these receptor proteins relative to normal meninges. In addition, the expression of phosphorylated/activated ErbB1/EGFR1 and phosphorylated/activated ErbB2/HER2 was significantly associated with histological malignancy grade and prognosis, respectively. The concurrent upregulation of ErbB receptors in human meningioma supports their fundamental role in the tumourigenesis of these tumours, and they could thus be exploited in diagnostics, prognosis, and ultimately, in targeted clinical interventions.

Decreased expression of ErbB2 on left ventricular epicardial cells in patients with diabetes mellitus

We investigated the cell surface expression of ErbB receptors on left ventricular (LV) epicardial endothelial cells and CD105+ cells obtained from cardiac biopsies of patients undergoing coronary artery bypass grafting surgery (CABG). Endothelial cells and CD105+ non-endothelial cells were freshly isolated from LV epicardial biopsies obtained from 15 subjects with diabetes mellitus (DM) and 8 controls. The expression of ErbB receptors was examined using flow cytometry. We found that diabetes mellitus (DM) and high levels of hemoglobin A1C are associated with reduced expression of ErbB2. To determine if the expression of ErbB2 receptors is regulated by glucose levels, we examined the effect of high Glucose in human microvascular endothelial cells (HMEC-1) and CD105+ non-endothelial cells, using a novel flow cytometric approach to simultaneously determine the total level, cell surface expression, and phosphorylation of ErbB2. Incubation of cells in the presence of 25 mM d-glucose resulted in decreased cell surface but not total levels of ErbB2. The level of ErbB2 at the cell surface is controlled by disintegrin and metalloproteinase domain-containing protein 10 (ADAM10) that is expressed on LV epicardial cells. Inhibition of ADAM10 prevented the high glucose-dependent decrease in the cell surface expression of ErbB2. We suggest that high Glucose depresses ErbB receptor signaling in endothelial cells and cardiac progenitor cells via the promotion of ADAM10-dependent cleavage of ErbB2 at the cell surface, thus contributing to vascular dysfunction and adverse remodeling seen in diabetic patients.

300 EGF System receptors status in aggressive subtypes of endometrial cancer

Introduction/Aims: ErbB receptors are trans-membrane glycoproteins with tyrosine kinase activity. Especially in cancer, they are implicated in cell proliferation, transformation, angiogenesis, migration and invasion. The expression of ErbB receptors, has not been studied well in endometrial cancer patients. The main aim of our study was to access EGF system receptors status in aggressive subtypes of endometrial cancer. Methods: We evaluated retrospectively tissue specimens from 10 patients with type II endometrial cancer, that have been treated in Gynaecological Oncology Unit of the University of Patras.

367 EGF System receptors profiling in young patients with cin 3 and cervical cancer

Introduction/Aims: ErbB receptors are trans-membrane glycoproteins with tyrosine kinase activity. Especially in cancer, they are implicated in cell proliferation, transformation, angiogenesis, migration and invasion. The expression of ErbB receptors, has not been studied well in patients with CIN 3 and cervical cancer. The main aim of our study was to access EGF system receptors status among young patients with CIN 3 and cervical cancer. Method: We evaluated retrospectively tissue specimens from 75 patients with CIN 3 and cervical cancer, that have been treated in Gynaecological Oncology Unit of the University of Patras.

299 EGF System receptors profiling in various histologic subgroups of endometrial cancer

Introduction/Aims: ErbB receptors are trans-membrane glycoproteins with tyrosine kinase activity. Especially in cancer, they are implicated in cell proliferation, transformation, angiogenesis, migration and invasion. The expression of ErbB receptors, has not been studied well in endometrial cancer patients. The main aim of our study was to assess EGF system receptors expression among various histologic subgroups of endometrial cancer. Methods: We evaluated retrospectively tissue specimens from 93 patients with endometrial cancer, that have been treated in Gynaecological Oncology Unit of the University of Patras.

Abstract 11303: Reduced Level of ErbB Receptors in Non-Myocyte Left Ventricular Cells in Patients with Diabetes Mellitus Undergoing Coronary Artery Bypass Grafting Surgery Contributes to Vascular Dysfunction

Introduction: ErbB signaling contributes to the regulation of vascular function through endothelial cell differentiation, proliferation, survival, and regulation of barrier integrity. Patients with diabetes (DM) are at an increased risk for endothelial dysfunction and worse outcomes after CABG. We investigated the level of ErbB expression on CD31 endothelial cells and CD105+ cells in DM and non-DM patients undergoing CABG. Methods: With IRB approval, left ventricular (LV) epicardial biopsies and blood samples from 25 patients (non-DM=9, DM=16), were obtained during surgery. Flow cytometry was used to determine the cell surface expression of ErbB receptors on cardiac endothelial and CD105+ cells isolated from the LV. Subpopulations of white blood cells were determined in blood samples. Plasma levels of IL-6, IL-8, and TNFα were measured. Results: Lower levels of ErbB1 and ErbB2 receptors expression on endothelial cells isolated from LV biopsies were found in patients with DM compared to non-DM control, and the expression of ErbB receptors negatively correlated with HgbA1c level. There were fewer CD105+ cells in DM patients (122 vs 153 cell/mg tissue, P=.021). Trends toward a negative association were found between the expression of ErbB2 in CD105+ cells and the number of neutrophils, as well as, HgbA1c level. No differences were found between DM and non-DM patients in the number of white blood cell subsets or plasma levels of IL-6, IL-8 and TNFα. Conclusions: We demonstrated that DM and high glucose blood levels are associated with reduced expression of ErbB1 and ErbB2 on endothelial cells and decreased number of CD105+ cells in the LV. Cardiac CD105+ are endothelial progenitor cells that differentiate in response to ErbB ligands. A decreased pool of CD105 endothelial progenitors in combination with down-regulation of ErbB1 and ErbB2 receptors in endothelial cells represent a novel mechanism for vascular dysfunction and adverse remodeling seen in diabetic patients.

Comprehensive analysis of the ErbB receptor family in pediatric nervous system tumors and rhabdomyosarcoma.

There is a paucity of knowledge regarding pediatric biomarkers, including the relevance of ErbB pathway aberrations in pediatric tumors. We investigated the occurrence of ErbB receptor aberrations across different pediatric malignancies, to identify patterns of ErbB dysregulation and define biomarkers suitable for patient enrichment in clinical studies. Tissue samples from 297 patients with nervous system tumors and rhabdomyosarcoma were analyzed for immunohistochemical expression or gene amplification of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2). Exploratory analyses of HER3/HER4 expression, and mRNA expression of ErbB receptors/ligands (NanoString) were performed. Assay validation followed general procedures, with additional validation to address Clinical Laboratory Improvement Amendments (CLIA) requirements. In most tumor types, samples with high ErbB receptor expression were found with heterogeneous distribution. We considered increased/aberrant ErbB pathway activation when greater than or equal to two EGFR/HER2 markers were simultaneously upregulated. ErbB pathway dysregulation was identified in ∼20%-30% of samples for most tumor types (medulloblastoma/primitive neuroectodermal tumors 31.1%, high-grade glioma 27.1%, neuroblastoma 22.7%, rhabdomyosarcoma 23.1%, ependymoma 18.8%), 4.2% of diffuse intrinsic pontine gliomas, and no recurrent or refractory low-grade astrocytomas. In medulloblastoma/primitive neuroectodermal tumors and neuroblastoma, this was attributed mainly to high EGFR polysomy/HER2 amplification, whereas EGFR gene amplification was observed in some high-grade glioma samples. EGFR/HER2 overexpression was most prevalent in ependymoma. Overexpression and/or amplification of EGFR/HER2 were identified as potential enrichment biomarkers for clinical trials of ErbB-targeted drugs.

Unique evolutionary trajectories of breast cancers with distinct genomic and spatial heterogeneity

Breast cancers exhibit intratumoral heterogeneity associated with disease progression and therapeutic resistance. To define the sources and the extent of heterogeneity, we performed an in-depth analysis of the genomic architecture of three chemoradiation-naïve breast cancers with well-defined clinical features including variable ER, PR, ERBB2 receptor expression and two distinct pathogenic BRCA2mut genotypes. The latter included a germ line carrier and a patient with a somatic variant. In each case we combined DNA content-based flow cytometry with whole exome sequencing and genome wide copy number variant (CNV) analysis of distinct populations sorted from multiple (4–18) mapped biopsies within the tumors and involved lymph nodes. Interrogating flow-sorted tumor populations from each biopsy provided an objective method to distinguish fixed and variable genomic lesions in each tumor. Notably we show that tumors exploit CNVs to fix mutations and deletions in distinct populations throughout each tumor. The identification of fixed genomic lesions that are shared or unique within each tumor, has broad implications for the study of tumor heterogeneity including the presence of tumor markers and therapeutic targets, and of candidate neoepitopes in breast and other solid tumors that can advance more effective treatment and clinical management of patients with disease.

Hypoxia-sensing CAR T cells provide safety and efficacy in treating solid tumors

SummaryUtilizing T cells expressing chimeric antigen receptors (CARs) to identify and attack solid tumors has proven challenging, in large part because of the lack of tumor-specific targets to direct CAR binding. Tumor selectivity is crucial because on-target, off-tumor activation of CAR T cells can result in potentially lethal toxicities. This study presents a stringent hypoxia-sensing CAR T cell system that achieves selective expression of a pan-ErbB-targeted CAR within a solid tumor, a microenvironment characterized by inadequate oxygen supply. Using murine xenograft models, we demonstrate that, despite widespread expression of ErbB receptors in healthy organs, the approach provides anti-tumor efficacy without off-tumor toxicity. This dynamic on/off oxygen-sensing safety switch has the potential to facilitate unlimited expansion of the CAR T cell target repertoire for treating solid malignancies.

LncRNA MIR503HG inhibits cell proliferation and promotes apoptosis in TNBC cells via the miR-224-5p/HOXA9 axis

Triple-negative breast cancer (TNBC) is a highly invasive subtype of breast cancer. This study investigated the molecular mechanism and influences of MIR503HG, miR-224-5p, and homeobox A9 (HOXA9) on TNBC cell growth and migration. Dual-luciferase reporter gene and RNA immunoprecipitation were performed to examine the regulation of MIR503HG, miR-224-5p, and HOXA9. Cell proliferation, apoptosis, migration, and invasion were evaluated by colony formation, flow cytometry, and Transwell assays. Finally, nude mice were employed to investigate the influence of MIR503HG on TNBC tumor growth. HOXA9 protein levels were detected by immunohistochemical staining. MIR503HG and HOXA9 expression were reduced in TNBC, while miR-224-5p was increased. Overexpression of MIR503HG or HOXA9 reduced the cell migration ability and proliferation and promoted apoptosis, and knockdown of MIR503HG or overexpression of miR-224-5p exhibited the opposite effects. Furthermore, MIR503HG promoted HOXA9 expression by inhibiting miR-224-5p. Overexpression of miR-224-5p reversed the effects of MIR503HG overexpression on TNBC cells, while overexpression of HOXA9 reversed the effect of MIR503HG knockdown. Additionally, an in vivo study proved that MIR503HG inhibited TNBC tumor growth via the miR-224-5p/HOXA9 axis. MIR503HG inhibited cell proliferation and promoted the apoptosis of TNBC cells via the miR-224-5p/HOXA9 axis, which may function as a novel target for the treatment of TNBC.

Effect of Combining EGFR Tyrosine Kinase Inhibitors and Cytotoxic Agents on Cholangiocarcinoma Cells.

PurposeThe potential of members of the epidermal growth factor receptor (ErbB) family as drug targets in cholangiocarcinoma (CCA) has not been extensively addressed. Although phase III clinical trials showed no survival benefits of erlotinib in patients with advanced CCA, the outcome of the standard-of-care chemotherapy treatment for CCA, gemcitabine/cisplatin, is discouraging so we determined the effect of other ErbB receptor inhibitors alone or in conjunction with chemotherapy in CCA cells.Materials and MethodsErbB receptor expression was determined in CCA patient tissues by immunohistochemistry and digital-droplet polymerase chain reaction, and in primary cells and cell lines by immunoblot. Effects on cell viability and cell cycle distribution of combination therapy using ErbB inhibitors with chemotherapeutic drugs was carried out in CCA cell lines. 3D culture of primary CCA cells was then adopted to evaluate the drug effect in a setting that more closely resembles in vivo cell environments.ResultsCCA tumors showed higher expression of all ErbB receptors compared with resection margins. Primary and CCA cell lines had variable expression of erbB receptors. CCA cell lines showed decreased cell viability when treated with chemotherapeutic drugs (gemcitabine and 5-fluorouracil) but also with ErbB inhibitors, particularly afatinib, and with a combination. Sequential treatment of gemcitabine with afatinib was particularly effective. Co-culture of CCA primary cells with cancer-associated fibroblasts decreased sensitivity to chemotherapies, but sensitized to afatinib.ConclusionAfatinib is a potential epidermal growth factor receptor targeted drug for CCA treatment and sequential treatment schedule of gemcitabine and afatinib could be explored in CCA patients.

Abstract A16: Characterization of ErbB family expression in intratibial bladder cancer xenografts: Considerations for preclinical modeling

Abstract Background: Approximately 40% of patients with distant metastatic bladder cancer are diagnosed with bone metastases and a dismal 5-year survival rate of less than 10%. The bone has long been thought to act as “soil” for metastatic cells to “seed,” likely due to the favorable, albeit complex microenvironment that comprises various cell types and growth factors. A study by The Cancer Genome Atlas (TCGA) profiling 405 muscle-invasive bladder cancers reported alterations of the receptor tyrosine kinase pathway in 20% of these patients. This included amplification of EGFR in 9% of cases, mutation or amplification of ERBB2 in 9% of cases, and mutation of ERBB3 in 6% of cases. The purpose of this study was to correlate the expression of ErbB family members in human bladder cancer cell lines with their ability to grow as intratibial xenografts in immune-compromised mice. Methods: We performed intratibial injections with six established human bladder cancer cell lines in 22 NSG mice and we allowed these xenografts to grow for five weeks. Four cell lines expressed luciferase and were measured for bioluminescent signals at multiple time points. The resulting formalin-fixed, paraffin-embedded tibiae were processed for immunohistochemistry (IHC) which utilized human specific antibodies to CK8, EGFR, HER2, Ki-67, and uroplakin. Results: The four luciferase-expressing cell lines exhibited increasing bioluminescent signal over the duration of the study (n=14). Five cell lines had positive human anti-Ki-67 staining in the tibia, confirming their growth in the bone after the intratibial injection. All five cell lines maintained expression, and in certain cases overexpression, of EGFR and HER2 in the intratibial xenografts. We observed an 87.5% take rate (n=16) in a subset of bladder cell lines (5 out of 6) and only the UM-UC-12 cell line exhibiting a 0% take rate (n=6). Conclusion: The high take rate of these five cell lines in the tibia exceeded that of previous studies (25-50%), indicating that the take rate is cell line dependent. Characterizing the ErbB expression profile of individual cell lines in the tibia xenograft model is an important consideration before using these xenografts as a preclinical model. Furthermore, ubiquitous expression of ErbB receptors in tibiae xenografts highlights the importance of growth factor signaling in metastatic bladder cancer and the potential for ErbB targeted therapy. Citation Format: Luke J. Broses, Kathleen C. Day, Aaron M. Udager, Mark L. Day. Characterization of ErbB family expression in intratibial bladder cancer xenografts: Considerations for preclinical modeling [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2019 May 18-21; Denver, CO. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(15_Suppl):Abstract nr A16.

Abstract 432: Loss of Primary Cilia PertErbBs Ventricular Maturation in the Developing Heart

The embryonic heart requires cardiac neural crest cells (CNCC), which elaborate the sensory organelle called the primary cilium and assist in the development of the cardiac valves, the interventricular septum, and the outflow tract. Our laboratory has modeled congenital heart defects (CHD) resulting from CNCC-specific ciliary loss, using a Wnt1:Cre-2, Ift88 -targeted conditional elimination of primary cilia. Ift88 -homozygous mutants (MUT) displayed a variety of predicted CNCC-associated CHDs, but also revealed a novel disorganization of the ventricular endocardium, pronounced noncompaction of the ventricular myocardium, and perinatal lethality. Neuregulin-1 / ErbB signaling is a well-known key modulator of myocardial development as well as the compaction process of the ventricular myocardium. Flow cytometric analysis of embryonic day 12.5 (E12.5) whole hearts revealed that loss of cilia in CNCC of MUT hearts led to a 43% reduction in cardiomyocytes (CM), defined as VCAM-1-positive/CD31-negative cells (p<0.01), and a trend towards increased CD31-positive endocardial cell (EC) populations (34% increase; p = 0.09) when compared to control (CON) hearts. Analysis of cell surface expression of ErbB receptors revealed a slight decrease in ErbB2 and ErbB4 expression in VCAM-1-positive CM of MUT with no change in ErbB3 expression, and decreased ErbB4 expression in MUT EC. Immunofluorescence analysis of proliferation was quantified in various cardiac subpopulations, including cardiac Troponin-T-positive (cTnT+/CM) cells in the ventricular myocardium. While overall cell density at E12.5 did not differ between the MUT and CON (p=0.43), CM proliferation increased significantly in MUT (5.50%) when compared to CON (2.70%; p<0.01). Taken together, decreased ErbB4 expression in the expanding MUT EC population, combined with decreased CM numbers, could serve as a novel mechanistic explanation for the pronounced hypertrabeculation and noncompaction of the MUT phenotype seen at E14.5. Our results indicate that loss of CNCC primary cilia leads to ventricular noncompaction, with a shift in the relative proportion and dynamics of both CM and EC subpopulations, which is potentially attributable to impaired ErbB signaling of the developing endocardium.

Triple-Negative Breast Cancer: A Review of Conventional and Advanced Therapeutic Strategies.

Triple-negative breast cancer (TNBC) cells are deficient in estrogen, progesterone and ERBB2 receptor expression, presenting a particularly challenging therapeutic target due to their highly invasive nature and relatively low response to therapeutics. There is an absence of specific treatment strategies for this tumor subgroup, and hence TNBC is managed with conventional therapeutics, often leading to systemic relapse. In terms of histology and transcription profile these cancers have similarities to BRCA-1-linked breast cancers, and it is hypothesized that BRCA1 pathway is non-functional in this type of breast cancer. In this review article, we discuss the different receptors expressed by TNBC as well as the diversity of different signaling pathways targeted by TNBC therapeutics, for example, Notch, Hedgehog, Wnt/b-Catenin as well as TGF-beta signaling pathways. Additionally, many epidermal growth factor receptor (EGFR), poly (ADP-ribose) polymerase (PARP) and mammalian target of rapamycin (mTOR) inhibitors effectively inhibit the TNBCs, but they face challenges of either resistance to drugs or relapse. The resistance of TNBC to conventional therapeutic agents has helped in the advancement of advanced TNBC therapeutic approaches including hyperthermia, photodynamic therapy, as well as nanomedicine-based targeted therapeutics of drugs, miRNA, siRNA, and aptamers, which will also be discussed. Artificial intelligence is another tool that is presented to enhance the diagnosis of TNBC.

Tumor cell sensitivity to vemurafenib can be predicted from protein expression in a BRAF-V600E basket trial setting

BackgroundGenetics-based basket trials have emerged to test targeted therapeutics across multiple cancer types. However, while vemurafenib is FDA-approved for BRAF-V600E melanomas, the non-melanoma basket trial was unsuccessful, suggesting mutation status is insufficient to predict response. We hypothesized that proteomic data would complement mutation status to identify vemurafenib-sensitive tumors and effective co-treatments for BRAF-V600E tumors with inherent resistance.MethodsReverse Phase Proteomic Array (RPPA, MD Anderson Cell Lines Project), RNAseq (Cancer Cell Line Encyclopedia) and vemurafenib sensitivity (Cancer Therapeutic Response Portal) data for BRAF-V600E cancer cell lines were curated. Linear and nonlinear regression models using RPPA protein or RNAseq were evaluated and compared based on their ability to predict BRAF-V600E cell line sensitivity (area under the dose response curve). Accuracies of all models were evaluated using hold-out testing. CausalPath software was used to identify protein-protein interaction networks that could explain differential protein expression in resistant cells. Human examination of features employed by the model, the identified protein interaction networks, and model simulation suggested anti-ErbB co-therapy would counter intrinsic resistance to vemurafenib. To validate this potential co-therapy, cell lines were treated with vemurafenib and dacomitinib (a pan-ErbB inhibitor) and the number of viable cells was measured.ResultsOrthogonal partial least squares (O-PLS) predicted vemurafenib sensitivity with greater accuracy in both melanoma and non-melanoma BRAF-V600E cell lines than other leading machine learning methods, specifically Random Forests, Support Vector Regression (linear and quadratic kernels) and LASSO-penalized regression. Additionally, use of transcriptomic in place of proteomic data weakened model performance. Model analysis revealed that resistant lines had elevated expression and activation of ErbB receptors, suggesting ErbB inhibition could improve vemurafenib response. As predicted, experimental evaluation of vemurafenib plus dacomitinb demonstrated improved efficacy relative to monotherapies.Conclusions: Combined, our results support that inclusion of proteomics can predict drug response and identify co-therapies in a basket setting.

Clinicopathological and prognostic values of ErbB receptor family amplification in primary osteosarcoma

Osteosarcoma is a malignant bone tumor with extremely high invasion, metastasis and mortality. The prognosis of patients with osteosarcoma remains poor. The ErbB receptor family was found to be overexpressed in human cancers and associated with poor prognosis. However, the role of ErbB receptor family in osteosarcoma has not been fully understood. The present study aimed to investigate the clinicopathological and prognostic significances of ErbB receptors in primary osteosarcoma. Western blot (WB), reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and fluorescence in situ hybridization (FISH) were used to detect the protein and gene expression of ErbB receptors in 60 primary osteosarcoma specimens and 30 non-neoplastic bone tissues. WB and RT-qPCR analyses showed that the protein and mRNA expression levels of EGFR, ErbB3 and ErbB4 in osteosarcoma specimens were significantly higher than those in non-neoplastic bone tissues. Seventeen (28.33%), 15 (25.00%) and 15 (25.00%) osteosarcoma specimens presented with amplification of EGFR, ErbB3 and ErbB4 gene, respectively, which were significantly higher compared with non-neoplastic bone tissues. The amplification of ErbB3 and ErbB4 in osteosarcoma was associated with advanced surgical stage. The amplification of EGFR, ErbB3, ErbB4 and the co-amplification of EGFR-ErbB3, EGFR-ErbB4, ErbB3-ErbB4 was linked with poor response to chemotherapy and distant metastasis. The amplification of EGFR, ErbB3 and ErbB4, as well as their co-amplification demonstrated independent prognostic values for reduced survival time of osteosarcoma patients and may serve as potential therapeutic targets for osteosarcoma patients in the future.

Receptor Tyrosine Kinase Signaling Networks Define Sensitivity to ERBB Inhibition and Stratify Kras-Mutant Lung Cancers.

Most non-small cell lung cancers (NSCLC) contain nontargetable mutations, including KRAS, TP53, or STK11/LKB1 alterations. By coupling ex vivo drug sensitivity profiling with in vivo drug response studies, we aimed to identify drug vulnerabilities for these NSCLC subtypes. Primary adenosquamous carcinoma (ASC) or adenocarcinoma (AC) cultures were established from KrasG12D/+;Lkb1fl/fl (KL) tumors or AC cultures from KrasG12D/+;p53fl/fl (KP) tumors. Although p53-null cells readily propagated as conventional cultures, Lkb1-null cells required conditional reprograming for establishment. Drug response profiling revealed short-term response to MEK inhibition, yet long-term clonogenic assays demonstrated resistance, associated with sustained or adaptive activation of receptor tyrosine kinases (RTK): activation of ERBBs in KL cultures, or FGFR in AC cultures. Furthermore, pan-ERBB inhibition reduced the clonogenicity of KL cultures, which was exacerbated by combinatorial MEK inhibition, whereas combinatorial MEK and FGFR inhibition suppressed clonogenicity of AC cultures. Importantly, in vivo studies confirmed KL-selective sensitivity to pan-ERBB inhibition, which correlated with high ERBB ligand expression and activation of ERBB receptors, implying that ERBB network activity may serve as a predictive biomarker of drug response. Interestingly, in human NSCLCs, phosphorylation of EGFR or ERBB3 was frequently detected in ASCs and squamous cell carcinomas. We conclude that analysis of in situ ERBB signaling networks in conjunction with ex vivo drug response profiling and biochemical dissection of adaptive RTK activities may serve as a valid diagnostic approach to identify tumors sensitive to ERBB network inhibition.