Supplementary Material for: Comprehensive DNA Methylation and Extensive Mutation Analyses of HER2-Positive Breast Cancer

<b><i>Objective:</i></b> Resistance to trastuzumab is a problem that remains to be solved in HER2-positive breast cancer. We aimed to characterize profiles of genetic and epigenetic alterations in cancer-related pathways in HER2-positive breast cancers, using biopsy tissue samples obtained from patients enrolled in a prospective neoadjuvant clinical trial. <b><i>Methods:</i></b> HER2-positive breast cancer tissue samples were collected and processed with the PAXgene Tissue System. A total of 24 breast cancers were analyzed. Genetic alterations of 409 cancer-related genes were analyzed by a bench-top next-generation sequencer. DNA methylation statuses were analyzed by a bead array with 485,512 probes. <b><i>Results:</i></b> The WNT pathway was potentially activated by aberrant methylation of its negative regulators, such as <i>DKK3</i> and <i>SFRP1</i>, in 9 breast cancers. The AKT/mTOR pathway was activated by mutations of <i>PIK3CA</i> in 5 breast cancers. The Notch pathway was potentially activated by mutations of <i>NOTCH1</i> and <i>NOTCH2</i> in 4 breast cancers. The p53 pathway was inactivated by mutations of <i>TP53</i> in 13 breast cancers and potentially by aberrant methylation of its downstream genes in 10 breast cancers. Cell adhesion was affected by mutations of <i>CDH1</i> in 1 breast cancer. <b><i>Conclusion:</i></b> Genes involved in cancer-related pathways were frequently affected not only by genetic but also by epigenetic alterations in HER2-positive breast cancer

Supplementary Material for: Contemplation of the effect of nivolumab plus cabosantinib therapy on cerebral hemorrhage in patients with brain metastasis of renal cell carcinoma: A case report

Although the response to combination therapy has been reported in patients with brain metastases from advanced renal cancer, treatment-related cerebral hemorrhage has not been adequately studied. The CheckMate 9ER clinical trial of nivolumab and cabozantinib excluded patients with brain metastases. Therefore, the associated treatment outcomes in these patients with brain metastases are unclear. Herein, we report a case of bleeding from brain metastases in a patient with advanced renal cancer after gamma knife combination therapy with nivolumab and cabozantinib. Fortunately, the cerebral hemorrhage of the patient was alleviated by conservative treatment. Despite treatment interruption, the metastatic lesions reduced in size, and treatment was gradually resumed. In this case study, we report the risk of cerebral hemorrhage in combination therapy for brain metastasis cases, how to manage hemorrhage cases, and their prognosis

Supplementary Material for: MicroRNA-372 Is Associated with Poor Prognosis in Colorectal Cancer

<i>Objective:</i> MicroRNA-372 <i>(miR-372)</i> is reportedly shown to be an oncogene in human testicular germ cell tumors and gastric cancers, but its expression in colorectal cancer (CRC) is not yet determined. This study investigated the clinical significance of <i>miR-372</i> expression in CRC. <i>Methods: </i>qRT-PCR was used to evaluate <i>miR-372</i> in 144 CRC patients, and large tumor suppressor 2 <i>(LATS2)</i> expression was also examined as the likely target gene of <i>miR-372</i>. In vitroassays were performed to evaluate the biological function of<i> miR-372. </i><i>Results: </i>Multivariate analysis indicated that high<i> miR-372</i> expression was an independent prognostic factor (p = 0.006). High <i>miR-372</i> expression was associated with synchronous liver metastasis (p = 0.035). We found an inverse relationship between<i> miR-372 </i>and <i>LATS2 by</i> qRT-PCR (p = 0.007) and immunohistochemistry (p = 0.042) using CRC tissue samples. Furthermore, pre-miR-372 led to a decrease in the LATS2 protein and an increase in proliferative activity of LoVo cells. We also found a significant association between low <i>LATS2</i> expression and liver metastasis (p = 0.042). <i>Conclusions:</i> This study suggested that <i>miR-372</i> was a novel independent prognostic factor in CRC. Our data suggest that <i>LATS2 </i>may serve as one of the target genes of <i>miR-372 </i>in clinical CRC tissues