Neu Gene Research Articles

Prognostic value of Her-2/neu and clinicopathologic factors for evaluating progression and disease-specific death in Chinese men with prostate cancer.

Her-2/neu gene overexpression has been found in several malignancies, and is associated with poor prognosis; while its role in the tumorigenesis and progression of prostate cancer (PCa) is still controversial. This study aimed to evaluate the prognostic value of Her-2/neu protein expression and clinicopathologic factors in antiandrogen-treated Chinese men with PCa for disease progression and PCa-specific death. Her-2/neu protein expression was determined using immunohistochemistry (IHC) in specimens collected from 124 prostate biopsies and transurethral resection of prostate (TURP) from seven prostate cancer patients. Her-2/neu protein expression was 0, 1+, 2+, and 3+ in 40 (30.5%), 8 (6.1%), 67 (51.1%), and 16 (12.2%) cases, respectively. Her-2/neu protein expression showed significant correlation as judged by Gleason score (P = 0.049), clinical tumor-node-metastases (cTNM) stage (P = 0.018) and disease progression (P = 0.001), but did not correlate with prostate-specific antigen (PSA) (P = 0.126) or PCa-specific death (P = 0.585). PSA (P = 0.001), Gleason score (P = 0.017), cTNM (P = 0.000) and Her-2/neu protein expression (P = 0.001) had prognostic value for evaluating the progression of PCa in univariate analysis. In Kaplan-Meier plots, both Gleason score (P = 0.035) and cTNM (P = 0.013) correlated with PCa-specific death. In multivariate analysis, only cTNM was significant for both disease progression (P = 0.001) and PCa-specific death (P = 0.031). Her-2/neu protein expression is significantly correlated with Gleason score, cTNM and disease progression, although it is not an independent predictor of disease progression and PCa-specific death. cTNM staging serves as an independent prognostic factor for disease progression and PCa-specific death.

Recent Advances in Comprehending the Signaling Pathways Involved in the Progression of Breast Cancer.

This review describes recent advances in the comprehension of signaling pathways involved in breast cancer progression. Calcium sensing receptor (CaSR), caveolae signaling, signaling referred to hypoxia-inducing factors and disturbances in the apoptotic machinery are related to more general biological mechanisms and are considered first. The others refer to signaling pathways of more specific biological mechanisms, namely the heparin/heparin-sulfate interactome, over-expression of miRNA-378a-5p, restriction of luminal and basal epithelial cells, fatty-acid synthesis, molecular pathways related to epithelial to mesenchimal transition (EMT), HER-2/neu gene amplification and protein expression, and the expression of other members of the epithelial growth factor receptor family. This progress in basic research is fundamental to foster the ongoing efforts that use the new genotyping technologies, and aim at defining new prognostic and predictive biomarkers for a better personalized management of breast cancer disease.

P.399 - Effects of long-term eteplirsen treatment on upper limb function in patients with Duchenne muscular dystrophy: findings of two phase 2 clinical trials

Duchenne muscular dystrophy (DMD) is a progressive, degenerative neuromuscular disease caused by gene mutations that result in the absence or deficiency of dystrophin protein. Absence of dystrophin compromises the regenerative ability of muscle fibers, and, as muscle deterioration overtakes muscle growth, loss of upper limb function causes difficulties in performing daily activities and negatively impacts quality of life. Eteplirsen (Exondys 51; Sarepta Therapeutics, Cambridge, MA), the first FDA approved treatment for DMD in the US, is indicated for treatment of patients with DMD amenable to exon 51 skipping. Eteplirsen has not been approved for marketing outside of the United States. We evaluated long-term upper limb function outcomes in patients aged 7–13 years with DMD amenable to exon 51 skipping who participated in 2 eteplirsen clinical trials. Study 201, a 28-week trial, randomized patients to receive once-weekly intravenous eteplirsen infusions (30 mg/kg, n = 4; 50 mg/kg, n = 4) or placebo (n = 4) for 24 weeks. At week 25, all patients received open-label eteplirsen through week 28. In study 202, an open-label trial, all patients received once-weekly eteplirsen at the same dose as in study 201 through 240 weeks. Upper limb function tests included the 9-Hole Peg Test (seconds), a maximum voluntary isometric contraction test (MVICT) with a Quantitative Movement Assessment system to assess elbow flexion and extension and hand grip (kg). Study 201 enrolled 12 patients; all completed extension study 202. Following confirmation of dystrophin production, patients showed relative stability or minimal decline of upper limb function on all measures assessed through week 240. In conclusion, eteplirsen may slow the deterioration of upper arm function in patients with DMD amenable to exon 51 skipping.

Use of Exfoliative Specimens and Fine-Needle Aspiration Smears for Mutation Testing in Lung Adenocarcinoma

Objective: Cytology specimens are considered to be a promising alternative for detecting driver mutations in lung cancer patients. We aimed to explore the suitability and utility of various cytology samples of non-small-cell lung cancer (NSCLC) patients for mutation testing. In addition to mutation detection, the importance of preanalytic factors was discussed. Design: A total of 116 cytology samples including 32 controls comprising pleural effusions, bronchial washings, and direct fine-needle aspiration (FNA) smears were included in the study for the detection of EGFR, KRAS, and Her-2/neu gene mutations. Tumor content was checked by microscopic evaluation. Tumor enrichment was done by scraping direct smears. DNA yield was assessed before selecting the method of mutation detection. Sanger sequencing and real-time PCR-based methods were used. Results: Overall, 20.23% EGFR mutations and 2.74% KRAS mutations were observed in this study. Nondriver genetic polymorphisms were observed in EGFR exon 20 in 37% cases. The coexistence of the EGFR mutation and EGFR polymorphism was seen in 7 cases. DNA yield was better in pleural effusions and bronchial washings. Real-time PCR was used in specimens of low DNA yield. Conclusions: Cytology samples are useful in ascertaining molecular diagnostic information for treatment purposes if they are optimized judiciously in their preanalytic phase.

Abstract 4807: Braf mutations initiate the development of rat gliomas induced by postnatal exposure to N-ethyl-N-nitrosourea (ENU)

Abstract Purpose A single dose of N-ethyl-N-nitrosourea (ENU) during late prenatal or early postnatal development induces a high incidence of malignant schwannomas and gliomas in rats. Although T -> A mutations in the transmembrane domain of the neu (c-ErbB-2) gene are the driver mutations in ENU-induced malignant schwannomas, the molecular basis of ENU-induced gliomas was unknown. The objectives of this study were to identify driver mutations in ENU-induced rat gliomas. Methods We performed whole-genome sequencing of gliomas that developed in three BDIV and two BDIX rats exposed to a single dose of 80 mg ENU/kg body weight on postnatal day one. Results T:A->A:T and T:A->C:G mutations, which are typical for ENU-induced mutagenesis, were predominant (41-55% of all somatic single nucleotide mutations). T->A mutations were detected in all 5 rat gliomas at Braf codon 545 (V545E), which corresponds to the human BRAF V600E. Additional screening revealed that 33 gliomas in BDIV rats and 12 gliomas in BDIX rats all carried a Braf V545E mutation, while peritumoral brain tissue of either strain (n=16) had the wild-type sequence. The gliomas were immunoreactive to BRAF V600E antibody. Conclusions Braf mutation is a frequent early event in the development of rat gliomas caused by a single dose of ENU. Citation Format: Kaishi Satomi, Qi Wang, Ji Eun Oh, Barbara Hutter, Benedikt Brors, Nicolle Diessl, Hai-Kun Liu, Stephan Wolf, Otmar Wiestler, Paul Kleihues, Bernd Koelsch, Andrea Kindler-Rohrborn, Hiroko Ohgaki. Braf mutations initiate the development of rat gliomas induced by postnatal exposure to N-ethyl-N-nitrosourea (ENU) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4807. doi:10.1158/1538-7445.AM2017-4807

Gilead buys Sarepta drug review voucher

Gilead Sciences has paid Sarepta Therapeutics $125 million for a voucher that can speed up U.S. regulatory review of a new drug application. The price tag, modest compared to past sales, has some observers wondering whether the program is losing its luster. Priority review vouchers (PRVs) were introduced in 2007 with the goal of spurring innovation in neglected tropical diseases. Expanded in 2012 to include rare childhood diseases, the program gives the developer of a drug with limited commercial potential a transferrable coupon that shaves four months off a future FDA drug review. Until Sarepta’s sale, the price of PRVs had generally been on the rise. Sanofi bought the first available PRV in 2014 for $67 million and later bought another one for $245 million. In 2015, AbbVie paid $350 million for its own fast-forward card. Gilead has bought three PRVs, one of which it used to accelerate the approval

Enrichment and Molecular Analysis of Breast Cancer Disseminated Tumor Cells from Bone Marrow Using Microfiltration

PurposeMolecular characterization of disseminated tumor cells (DTCs) in the bone marrow (BM) of breast cancer (BC) patients has been hindered by their rarity. To enrich for these cells using an antigen-independent methodology, we have evaluated a size-based microfiltration device in combination with several downstream biomarker assays.MethodsBM aspirates were collected from healthy volunteers or BC patients. Healthy BM was mixed with a specified number of BC cells to calculate recovery and fold enrichment by microfiltration. Specimens were pre-filtered using a 70 μm mesh sieve and the effluent filtered through CellSieve microfilters. Captured cells were analyzed by immunocytochemistry (ICC), FISH for HER-2/neu gene amplification status, and RNA in situ hybridization (RISH). Cells eluted from the filter were used for RNA isolation and subsequent qRT-PCR analysis for DTC biomarker gene expression.ResultsFiltering an average of 14×106 nucleated BM cells yielded approximately 17–21×103 residual BM cells. In the BC cell spiking experiments, an average of 87% (range 84–92%) of tumor cells were recovered with approximately 170- to 400-fold enrichment. Captured BC cells from patients co-stained for cytokeratin and EpCAM, but not CD45 by ICC. RNA yields from 4 ml of patient BM after filtration averaged 135ng per 10 million BM cells filtered with an average RNA Integrity Number (RIN) of 5.3. DTC-associated gene expression was detected by both qRT-PCR and RISH in filtered spiked or BC patient specimens but, not in control filtered normal BM.ConclusionsWe have tested a microfiltration technique for enrichment of BM DTCs. DTC capture efficiency was shown to range from 84.3% to 92.1% with up to 400-fold enrichment using model BC cell lines. In patients, recovered DTCs can be identified and distinguished from normal BM cells using multiple antibody-, DNA-, and RNA-based biomarker assays.

Crime against Humanity<br/>—Uncovering Two Decades of Corruption in the FDA Regarding DMD Treatment

The world needs to know the facts as to how the FDA “protects” the public health by assuring the safety, effectiveness, and security of human drugs and biological products for human use. Recent uproar has surfaced over the FDA’s approval process on the unproven Duchenne Muscular Dystrophy (DMD) drug, Exondys 51 (Sarepta Therapeutics, Cambridge, MA, USA), but what many people do not know are the unlawful events that occurred nearly two decades ago on a proven and promising treatment for DMD.

Fluorescent in Situ Hybridization and Real-Time Quantitative Polymerase Chain Reaction to Evaluate HER-2/neu Status in Breast Cancer

Breast cancer remains the most common and second lethal cancer in females. HER-2/neu is one of the most important amplified oncogene in breast cancer. The amplification of HER-2 is correlated with decreased survival, metastasis, and early recurrence. The amplification of HER-2/neu gene and synthesis of the protein are reported in 10%-34% of breast cancer cases associated with tumor size, advanced tumor stage, high-grade tumor, young age at diagnosis, absence of steroid hormone receptor, and lymph node involvement. Fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) methods are options to evaluate HER-2 expression. The current study aimed at identifying the correlation between FISH and real-time polymerase chain reaction (PCR) in measuring HER-2 expression. The study investigated the performance of the real-time PCR as measured against FISH method in IHC +2 borderline cases. In a total of 120 IHC 2+ samples, 58.3% were negative and 41.6% positive for HER-2 gene, confirmed by FISH as a gold standard method. The real-time PCR ratio was <1.8 for a majority (82.8%) of the tumor samples with unamplified HER-2 gene by FISH as a gold standard assay. Despite the fact that real-time PCR is a promising method to evaluate HER-2 over expression and a supplementary array to FISH, according to the results of the present study it cannot be utilized instead of gold standard techniques; therefore, additional studies should be carried out to appraise the value of this method to evaluate HER-2 over expression.

HER2/neu Testing In 432 Consecutive Breast Cancer Cases using FISH and IHC - A Comparative Study.

The development of trastuzumab, a HER-2/neu targeted monoclonal antibody resulted in significant improvements in clinical response and survival in HER-2/neu gene amplified group of patients. Thus, accurate assessment of HER-2/neu status becomes critical. Fluorescence In Situ Hybridization (FISH) and Immunohistochemistry (IHC) are the most commonly used methods for this purpose and specific recommendations exist with regard to the concordance to be observed between the two tests. Here, we report and evaluate the concordance rate between FISH and IHC for HER-2/neu status in breast cancer specimens. Archival paraffin blocks of tumour tissue from 450 patients of breast cancer were analyzed for Her-2/neu status using FISH and IHC. There was a highly significant concordance between the results of FISH and IHC assays in HER-2/neu status assessment in invasive breast cancer cases. There were inverse associations between the expression of Oestrogen Receptors/Progesterone Receptors (ER/PR) and HER-2/neu amplification. Although, IHC gave significant concordant results with FISH in determining HER-2/neu status, its subjective grading system precludes its use as a gold standard. FISH should always be used to determine true gene amplification when the IHC results are equivocal.

Eteplirsen: First Global Approval.

Eteplirsen (Exondys51) is an antisense oligonucleotide designed to induce exon 51 skipping that is developed by Sarepta Therapeutics. Intravenous eteplirsen has received accelerated approval from the US FDA for the treatment of Duchenne muscular dystrophy (DMD) in patients with a confirmed mutation of the DMD gene amenable to exon 51 skipping. Eteplirsen has orphan drug designation in the USA and EU, and rare paediatric disease designation in the USA for use in DMD. In the phaseIII PROMOVI trial, eteplirsen significantly increased dystrophin levels from baseline in muscle tissues of 12 evaluable patients with DMD after 48 weeks of treatment. This finding is supported by data from phase II trials. Long-term treatment with eteplirsen was associated with a decrease in the rate of decline in ambulation and pulmonary function in an open-label extension of a phase II trial. Eteplirsen was generally well tolerated in clinical trials. This article summarizes the milestones in the development of eteplirsen leading to this first approval for DMD.

Eteplirsen approved for Duchenne muscular dystrophy.

FDA on September 19 approved eteplirsen injection, an antisense oligonucleotide therapy marketed by Sarepta Therapeutics, for the treatment of patients with Duchenne muscular dystrophy (DMD) who have a confirmed mutation of the dystrophin gene that renders it amenable to exon 51 skipping. Sarepta

Sarepta licenses Summit’s DMD drugs

In its second partnership since FDA approval last month of eteplirsen, its exon-skipping Duchenne muscular dystrophy treatment, Sarepta Therapeutics has licensed the European rights to Summit Therapeutics’ utrophin modulator pipeline. The deal includes ezutromid, which is in a Phase II study for treating DMD. Summit will get $40 million up front and could reap another $522 million in milestone payments. Late last month, Sarepta teamed with Catabasis to study combining its exon-skipping drugs with Catabasis’s NF-κB inhibitors.

FDA okays first DMD drug

Sarepta Therapeutics has won FDA approval of Exondys 51 (eteplirsen), the first drug sanctioned for treating Duchenne muscular dystrophy. Afflicting one in every 3,500 to 5,000 boys, DMD is a fatal genetic disorder characterized by progressive muscle deterioration. Approved under an accelerated program for drugs that treat life-threatening diseases, Exondys 51 is being made available on the basis of initial clinical data. “We eagerly await learning more about the efficacy of this drug through a confirmatory clinical trial that the company must conduct,” says Janet Woodcock, director of FDA’s Center for Drug Evaluation & Research. DMD is caused by mutations in the gene encoding for dystrophin, a protein that keeps muscle cells intact. Sarepta’s phosphorodiamidate morpholino oligomers prevent the reading of faulty gene regions during mRNA processing. This “exon skipping” method enables production of a truncated, but still functional, protein. Up to 80% of DMD patients have genotypes open to

Braf Mutations Initiate the Development of Rat Gliomas Induced by Postnatal Exposure to N-Ethyl-N-Nitrosourea

A single dose of N-ethyl-N-nitrosourea (ENU) during late prenatal or early postnatal development induces a high incidence of malignant schwannomas and gliomas in rats. Although T->A mutations in the transmembrane domain of the Neu (c-ErbB-2) gene are the driver mutations in ENU-induced malignant schwannomas, the molecular basis of ENU-induced gliomas remains enigmatic. We performed whole-genome sequencing of gliomas that developed in three BDIV and two BDIX rats exposed to a single dose of 80 mg ENU/kg body weight on postnatal day one. T:A->A:T and T:A->C:G mutations, which are typical for ENU-induced mutagenesis, were predominant (41% to 55% of all somatic single nucleotide mutations). T->A mutations were identified in all five rat gliomas at Braf codon 545 (V545E), which corresponds to the human BRAF V600E. Additional screening revealed that 33 gliomas in BDIV rats and 12 gliomas in BDIX rats all carried a Braf V545E mutation, whereas peritumoral brain tissue of either strain had the wild-type sequence. The gliomas were immunoreactive to BRAF V600E antibody. These results indicate that Braf mutation is a frequent early event in the development of rat gliomas caused by a single dose of ENU.

Assessment of HER-2/neu gene amplification and expression as a biomarker for radiotherapy and hormonal treated breast cancer patients in Upper Egypt

Assessment of HER-2/neu gene amplification and expression as a biomarker for radiotherapy and hormonal treated breast cancer patients in Upper Egypt

A Study of Evaluation of Immunoreactivity of c-erbB-2 and Histopathological Parameter in Esophageal Carcinoma

Abstrct Background: Squamous cell carcinoma is commonest histologic variant of carcinoma of esophagus (ESCC). It is predominant in developing countries and usually has a poor prognosis, because of its advanced stage at the time of diagnosis. Understanding the prognostic factors helps in selection of the most appropriate surgical approach in order to select patient for target directed therapies. HER-2/neu (C-erbB-2), member of human epidermal growth factor receptors (EGFR) is important, as it plays a crucial role in therapeutic implications of esophageal carcinoma. Considering the importance of c-erbB-2 expression and its prognostic significance in esophageal cancer, this study was undertaken. Result: Fifty cases of esophageal carcinoma including squamous cell carcinoma and adenocarcinoma studied showed M: F ratio of 1.9:1 with age ranging from 32 years to 80 years (mean age 59.1 years). There were 43(86%) cases of Squamous cell carcinoma with 65.1% positivity of which 10 (20%) cases were poorly diffentiated with a positivity of 60% ,20 (40%) cases were moderately diffentiated with a positivity of 65% and 09 (18%) were well differentiated with a positivity of 55.6% for c-erbB-2 staining. Seven (14%) cases of adenocarcinoma with a positivity of 85.7% and four (08%) cases of endoscopic biopsy have a positivity of 75% for c-erbB-2 staining. Conclusion: Present study results illustrate that HER-2/neu(c-erbB-2) gene amplification is a influential prognostic indicator in ESCC. Therefore, additional studies with more cases including supplementary techniques are compelling to verify molecular alterations involved in tumor progression, thus assisting in the advancement of modern therapies for ESCC.

Chromosome aberrations [dup(1q)] in endometrial cancer: Gene analysis of 54 surgical specimens in Turkey

ObjectiveWe aimed to evaluate the frequency of chromosomal aberrations and mutations in the k-ras or Her-2/neu genes in surgical specimens of endometrial carcinoma and their association with clinicopathological findings. Materials and methodsFifty-four patients who were treated for endometrial cancer between April 2010 and May 2011 at the Kocaeli University Obstetrics and Gynecology Department, Kocaeli, Turkey were enrolled in a prospective study. Clinical and histopathological findings were recorded. Genetic analysis, which included the detection of chromosomal deletions and duplications, as well as k-ras and Her-2/neu mutations, was performed on endometrial samples from surgical specimens. ResultsIn 70% of cases, tumor size was >2 cm or covered the entire uterine cavity, affecting mostly corpus (76%) and invading less than half of the myometrium (80%). Forty-six cases (86%) had endometrioid-type carcinoma, and early stage (Stage I, 65%) and higher grade (Grade II–III, 66%) tumors were predominant. Lymph node and lymphovascular involvement was positive in 11% and 28% of the patients, respectively. Chromosomal aberrations (deletion or duplication) and Her-2/neu and k-ras mutations were encountered in 44%, 15%, and 13% of surgical specimens, respectively. The most common chromosomal aberration was dup(1q) (n = 16). Oncogenic mutations in Her-2/neu or k-ras had no association with the severity of endometrial cancer, but the presence of chromosomal aberrations, as a whole or dup(1q) alone, were associated with higher tumor size, deeper myometrial invasion, advanced stage or grade, lymphovascular invasion, and lymph node involvement (p < 0.05 for all). ConclusionChromosomal aberrations, particularly dup(1q), are related to advanced disease in endometrial cancer. Genetic analysis of cancer tissues may provide important insights in determining disease prognosis.

HER-2/neu en cáncer de mama localmente avanzado

IntroductionBreast cancer is the first cause of death due to malignancies in women over 25 years old, with 70% of the tumours being locally advanced. The therapeutic decision is based on lymph node activity, receptor status, size of the primary tumour, and the clinical stage at the time of diagnosis. A new predictor is the HER-2/neu gene, which has emerged as a prognostic marker and therapeutic target. Literature reports an expression range from 9% to 74%, with an average relative risk of 2.74 for overall survival. ObjectiveTo determine the prevalence of HER-2/neu gene in patients with locally advanced breast cancer. Material and methodsA retrospective study, historical cohort was conducted on females older than 18 years of age with the diagnosis of locally advanced breast cancer, who were attended from June 2008 to January 2014. They had to have immunohistochemistry performed for hormonal receptors and positivity for HER-2/neu, as well as a known site of metastasis. ResultsOf the 182 files included, an expression of 19.7% of HER-2/neu was found, with a tumour recurrence in 36%. The major site was bone, with 30.7%, of which 15.3% had a recurrence within 12-24 months, followed by lung and multiple sites in 23%, central nervous system in 15%, and locoregional in 7.7%. ConclusionsThe frequency of HER-2/neu in locally advanced breast cancer was 19.7%, with a major site of relapse to bone. There was also an increased occurrence with lobular histology, the type of receptor status, and menopause.

Duchenne muscular dystrophy drugs face tough path to approval.

Highly anticipated as new disease-modifying treatments for Duchenne muscular dystrophy (DMD), therapeutics by BioMarin Pharmaceutical (Kyndrisa™; drisapersen) and Sarepta Therapeutics (eteplirsen; AVI-4658) both recently received negative FDA reviews and are now facing battles for approval in the U.S. At present, BioMarin is committed to working with the FDA to forge a pathway to approval following the failure of its NDA, while Sarepta awaits the formal decision on its NDA, which is expected by late May 2016. Despite the critical nature of both reviews, analysts consider that there is still a narrow possibility of approval of both drugs. According to Consensus forecasts from Thomson Reuters Cortellis for Competitive Intelligence, Kyndrisa is forecast to achieve sales of USD 533.71 million in 2021.