ABSTRACT
Efficacious therapeutic approaches are urgently needed to improve outcomes in patients with oesophageal adenocarcinoma (OAC). However, oncogenic drivers amenable to targeted therapy are limited and their functional characterisation is essential. Among few targeted therapies available, anti-human epidermal growth factor receptor 2 (HER2) therapy showed only modest benefit for patients with OAC. Herein, we investigated the potential oncogenic role of growth factor receptor bound protein 7 (GRB7), which is reported to be co-amplified with HER2 (ERBB2) in OAC. GRB7 was highly expressed in 15% of OAC tumours, not all of which could be explained by co-amplification with HER2, and was associated with a trend for poorer overall survival. Knockdown of GRB7 decreased proliferation and clonogenic survival, and induced apoptosis. Reverse phase protein array (RPPA) analyses revealed a role for PI3K, mammalian target of rapamycin (mTOR), MAPK, and receptor tyrosine kinase signalling in the oncogenic action of GRB7. Furthermore, the GRB7 and HER2 high-expressing OAC cell line Eso26 showed reduced cell proliferation upon GRB7 knockdown but was insensitive to HER2 inhibition by trastuzumab. Consistent with this, GRB7 knockdown in vivo with an inducible shRNA significantly inhibited tumour growth in cell line xenografts. HER2 expression did not predict sensitivity to trastuzumab, with Eso26 xenografts remaining refractory to trastuzumab treatment. Taken together, our study provides strong evidence for an oncogenic role for GRB7 in OAC and suggests that targeting GRB7 may be a potential therapeutic strategy for this cancer. © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons, Ltd. on behalf of The Pathological Society of Great Britain and Ireland.
Subject(s)
Adenocarcinoma/metabolism , Biomarkers, Tumor/metabolism , Esophageal Neoplasms/metabolism , GRB7 Adaptor Protein/metabolism , Adenocarcinoma/drug therapy , Adenocarcinoma/mortality , Adenocarcinoma/pathology , Animals , Antineoplastic Agents, Immunological/therapeutic use , Blotting, Western , Cell Line, Tumor , Esophageal Neoplasms/drug therapy , Esophageal Neoplasms/mortality , Esophageal Neoplasms/pathology , Female , Gene Knockdown Techniques , Humans , Immunohistochemistry , Mice , Mice, Inbred NOD , Mice, SCID , Neoplasm Transplantation , Prognosis , Receptor, ErbB-2/metabolism , Survival Analysis , Trastuzumab/therapeutic useABSTRACT
BACKGROUND: The cornea is a highly transparent structure covering the anterior one-fifth of the eyeball. The suitability of post-mortem donor corneas for keratoplasty is currently qualitatively assessed. This makes inferences prone to bias and subjective variability. This study aimed to develop a simple, feasible and cost-effective method to quantify corneal transparency. METHODS: An artificial anterior chamber was modified to provide a central transparent passage and a standardised pressure segment. All corneas graded 'fair' were included in this study. The corneoscleral buttons were mounted on the modified artificial anterior chamber. The mounted chamber was held in a horizontal position at a fixed distance from a white projection screen. The laser source was placed in alignment with an artificial anterior chamber so that it passed through the centre of the cornea. A camera mounted on a tripod stand was placed at a prefixed distance. An image of the scattered laser spot that formed after the laser passed through the mounted cornea on the screen was captured with a single digital camera and standardised settings. Image analysis was performed using ImageJ, an open platform for scientific image analysis. The average red pixel intensity, max intensity, and full-width half max were calculated. RESULTS: The average red intensity was 132.45 ± 6.65 SD. The mean for max intensity was 51.1 ± 3.78 SD and the full-width half max 787.7 ± 84.7 SD. CONCLUSION: Laser quantification is a simple and cost-effective method of quantifying corneal transparency. The study lends proof to the principle involved.
Subject(s)
Cornea/physiology , Diagnostic Techniques, Ophthalmological , Image Processing, Computer-Assisted/methods , Lasers , Postmortem Changes , Tissue Donors/classification , Tissue and Organ Harvesting/classification , Aged , Female , Humans , Light , Male , Middle Aged , Scattering, RadiationABSTRACT
Genetic alterations that potentiate PI3K signaling are frequent in prostate cancer, yet how different genetic drivers of the PI3K cascade contribute to prostate cancer is unclear. Here, we report PIK3CA mutation/amplification correlates with poor survival of patients with prostate cancer. To interrogate the requirement of different PI3K genetic drivers in prostate cancer, we employed a genetic approach to mutate Pik3ca in mouse prostate epithelium. We show Pik3caH1047R mutation causes p110α-dependent invasive prostate carcinoma in vivo Furthermore, we report that PIK3CA mutation and PTEN loss coexist in patients with prostate cancer and can cooperate in vivo to accelerate disease progression via AKT-mTORC1/2 hyperactivation. Contrasting single mutants that slowly acquire castration-resistant prostate cancer (CRPC), concomitant Pik3ca mutation and Pten loss caused de novo CRPC. Thus, Pik3ca mutation and Pten deletion are not functionally redundant. Our findings indicate that PIK3CA mutation is an attractive prognostic indicator for prostate cancer that may cooperate with PTEN loss to facilitate CRPC in patients.Significance: We show PIK3CA mutation correlates with poor prostate cancer prognosis and causes prostate cancer in mice. Moreover, PIK3CA mutation and PTEN loss coexist in prostate cancer and can cooperate in vivo to accelerate tumorigenesis and facilitate CRPC. Delineating this synergistic relationship may present new therapeutic/prognostic approaches to overcome castration/PI3K-AKT-mTORC1/2 inhibitor resistance. Cancer Discov; 8(6); 764-79. ©2018 AACR.See related commentary by Triscott and Rubin, p. 682This article is highlighted in the In This Issue feature, p. 663.
Subject(s)
Class I Phosphatidylinositol 3-Kinases/genetics , Mutation , PTEN Phosphohydrolase/genetics , Prostatic Neoplasms, Castration-Resistant/genetics , Animals , Cell Line, Tumor , Disease Progression , Gene Amplification , Gene Deletion , Humans , Male , Mice , Neoplasm Invasiveness , Neoplasms, Experimental , Prognosis , Survival AnalysisABSTRACT
Traumatic insults to the spinal cord induce both immediate mechanical damage and subsequent tissue degeneration leading to a substantial physiological, biochemical, and functional reorganization of the spinal cord. Various spinal cord injury (SCI) models have shown the adaptive potential of the spinal cord and its limitations in the case of total or partial absence of supraspinal influence. Meaningful recovery of function after SCI will most likely result from a combination of therapeutic strategies, including neural tissue transplants, exogenous neurotrophic factors, elimination of inhibitory molecules, functional sensorimotor training, and/or electrical stimulation of paralyzed muscles or spinal circuits. Peripheral nerve grafts provide a growth-permissive substratum and local neurotrophic factors to enhance the regenerative effort of axotomized neurons when grafted into the site of injury. Regenerating axons can be directed via the peripheral nerve graft toward an appropriate target, but they fail to extend beyond the distal graft-host interface because of the deposition of growth inhibitors at the site of SCI. One method to facilitate the emergence of axons from a graft into the spinal cord is to digest the chondroitin sulfate proteoglycans that are associated with a glial scar. Importantly, regenerating axons that do exit the graft are capable of forming functional synaptic contacts. These results have been demonstrated in acute injury models in rats and cats and after a chronic injury in rats and have important implications for our continuing efforts to promote structural and functional repair after SCI.
