Suraj Manjunath

Unicystic ameloblastoma of the mandible - an unusual case report and review of literature

Ameloblastoma is a true neoplasm of odontogenic epithelial origin. It is the second most common odontogenic neoplasm, and only odontoma outnumbers it in reported frequency of occurrence. Its incidence, combined with its clinical behavior, makes ameloblastoma the most significant odontogenic neoplasm. Unicystic ameloblastoma (UA) refers to those cystic lesions that show clinical, radiographic, or gross features of a mandibular cyst, but on histologic examination show a typical ameloblastomatous epithelium lining part of the cyst cavity, with or without luminal and/or mural tumor growth. It accounts for 5-15% of all intraosseous ameloblastomas. We report a case of unicystic ameloblastoma in a 30-year-old female, and review the literature.

A Majority of Low (1-10%) ER Positive Breast Cancers Behave Like Hormone Receptor Negative Tumors

Background: The 2010 guidelines by ASCO-CAP have mandated that breast cancer specimens with ≥1% positively staining cells by immunohistochemistry should be considered Estrogen Receptor (ER) positive. This has led to a subclass of low-ER positive (1-10%) breast cancers. We have examined the biology and clinical behavior of these low ER staining tumors. Methods: We have developed a probabilistic score of the “ER-positivity” by quantitative estimation of ER related gene transcripts from FFPE specimens. Immunohistochemistry for ER was done on 240 surgically excised tumors of primary breast cancer. Relative transcript abundance of 3 house-keeping genes and 6 ER related genes were determined by q-RT PCR. A logistic regression model using 3 ER associated genes provided the best probability function, and a cut-off value was derived by ROC analysis. 144 high ER (>10%), 75 ER negative and 21 low-ER (1-10%) tumors were evaluated using the probability score and the disease specific survival was compared. Results: Half of the low-ER positive tumors were assigned to the ER negative group based on the probability score; in contrast 95% of ER negative and 92% of the high ER positive tumors were assigned to the appropriate ER group (p<0.0001). The survival of the low-ER group was intermediate between that of the high ER positive and ER negative groups (p<0.05). Conclusion: Our results suggest that the newly lowered ASCO-CAP criteria for ER positivity, leads to the false categorization of biologically ER negative tumors as ER positive ones. This may have particular relevance to India, where we have a much higher proportion of ER negative tumors in general.

The epigenetic silencing of the estrogen receptor (ER) by hypermethylation of the ESR1 promoter is seen predominantly in triple-negative breast cancers in Indian women

The proportion of estrogen receptor (ER)-negative and triple-negative (TN) breast cancer in Indian women is higher than that reported in the West, and this difference persists even after their migration to the West. The causes for this significant difference are not entirely clear. Hypermethylation of the ER promoter, an epigenetic alteration, is known to be one of the mechanisms by which the expression of ER is suppressed. Two thirds of breast cancer specimens from an Indian center tested, using the highly sensitive, methylation-specific polymerase chain reaction (MSP) technique, were reported positive. We have used a quantitative assay, the MethyLight, to better assess the extent of methylation in the ESR1 promoter region in 98 breast cancer tumor specimens from Indian women. In addition, the amount of ER transcripts was determined by quantitative reverse transcriptase polymerase chain reaction. Using the stringent cutoff of at least 4% of the target sequence being methylated, 27% of TN tumors were methylated. In addition they demonstrated the highest levels of methylation. In contrast less than 2% ER-positive tumors were hypermethylated. While the proportion of hypermethylated tumors are lower in this study than that estimated using MSP, our results support the notion of increased epigenetic deregulations in ER-negative tumors in general and TN tumors in particular. The development of this assay also permits a rational approach to the selection of patients for clinical trials examining the efficacy of demethylating agents in the treatment of ER-negative breast cancer.

Estrogen Receptor Negative Breast Cancer in India: Do We Really Have Higher Burden of this Subtype?

ER negative and Triple negative breast cancers carry a poorer prognosis and are not amenable to hormone therapy. It has been previously observed that Indian patients with breast cancer have a higher tendency to have these tumours. Whether this is due to inherent biological differences in the tumours of our patients is a matter of much debate. We have analysed 250 patients of breast cancer for hormone receptor status, compared them with western series, and attempted to support the hypothesis that the higher ER negativity and triple negativity is indeed due to different tumour biology.

High expression of integrin β6 in association with the Rho–Rac pathway identifies a poor prognostic subgroup within HER2 amplified breast cancers

Integrin αvβ6 is involved in the transition from ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) of the breast. In addition, integrin β6 (ITGB6) is of prognostic value in invasive breast cancers, particularly in HER2+ subtype. However, pathways mediating the activity of integrin αvβ6 in clinical progression of invasive breast cancers need further elucidation. We have examined human breast cancer specimens (N = 460) for the expression of integrin β6 (ITGB6) mRNA by qPCR. In addition, we have examined a subset (N = 147) for the expression of αvβ6 integrin by immunohistochemistry (IHC). The expression levels of members of Rho–Rac pathway including downstream genes (ACTR2, ACTR3) and effector proteinases (MMP9, MMP15) were estimated by qPCR in the HER2+ subset (N = 59). There is a significant increase in the mean expression of ITGB6 in HER2+ tumors compared to HR+HER2‐ and triple negative (TNBC) subtypes (P = 0.00). HER2+ tumors with the highest levels (top quartile) of ITGB6 have significantly elevated levels of all the genes of the Rho–Rac pathway (P‐values from 0.01 to 0.0001). Patients in this group have a significantly shorter disease‐free survival compared to the group with lower ITGB6 levels (HR = 2.9 (0.9–8.9), P = 0.05). The mean level of ITGB6 expression is increased further in lymph node‐positive tumors. The increased regional and distant metastasis observed in HER2+ tumors with high levels of ITGB6 might be mediated by the canonical Rho–Rac pathway through increased expression of MMP9 and MMP15.

Identification of BRCA1 Deficiency Using Multi-Analyte Estimation of BRCA1 and Its Repressors in FFPE Tumor Samples from Patients with Triple Negative Breast Cancer

Purpose Apart from germ-line BRCA1-mutated breast cancers, a significant proportion of women with sporadic triple negative breast cancer (TNBC) sub-type are known to harbour varying levels of BRCA1-dysfuction. There is currently no established diagnostic method to identify these patients. Methods The analysis was performed on 183 primary breast cancer tumor specimens from our longitudinal case-series archived as formalin-fixed-paraffin-embedded (FFPE) blocks comprising 71 TNBCs and 112 Hormone receptor positive HER2 negative (HR+HER2-) tumors. Transcript levels of BRCA1 and two of its repressors ID4 and microRNA182 were determined by TaqMan quantitative PCR. BRCA1 protein was detected immunohistochemically with the MS110 antibody. Results The representation of BRCA1 and its repressor ID4 as a ratio led to improved separation of TNBCs from HR+HER2- compared to either measure by itself. We then dichotomised the continuous distribution of each of the three measurements (Protein, MIRNA and transcript:repressor ratio) into categories of deficient (0) and adequate (1). A composite BRCA1 Deficiency Score (BDS) was computed by the addition of the score for all three measures. Samples deficient on 2 or more measures were deemed to be BRCA1 deficient; and 40% of all TNBCs met this criterion. Conclusion We propose here a simple multi-level assay of BRCA1 deficiency using the BRCA1:ID4 ratio as a critical parameter that can be performed on FFPE samples in clinical laboratories by the estimation of only 3 bio-markers. The ease of testing will hopefully encourage adoption and clinical validation.

“Peripherally Inserted Central Catheters: Our Experience from a Cancer Research Centre”

Peripherally inserted central catheter (PICC) is a modern drug delivery system utilised in oncology practice. The purpose of this study was to determine the complications associated with PICCs within a one year study period. PICCs inserted in patients registered at Dharamshila Cancer Hospital and research centre from 1st July 2012 to 30th June 2013 were studied. Data was retrieved from the procedure room records, medical records department, department of radiology and department of microbiology. Data was collected by oncology residents and procedure team. A total of 246 PICCs were inserted during the one year period. Complete data was not available in 23 patients. 223 results were included in the final analysis. USG guidance was required in 14 patients (6.3%). Optimal PICC duration was achieved in 151 patients (67.7%). 28 patient developed culture positive infective complications (12.5%). 44 patients developed mechanical complications (19.7%). Our study shows a relatively higher rate of infective complications. PICC is an acceptable means of drug delivery system.

HR+HER2− breast cancers with growth factor receptor–mediated EMT have a poor prognosis and lapatinib downregulates EMT in MCF-7 cells

Despite an overall good prognosis, a significant proportion of patients with hormone receptor positive human epidermal growth factor receptor 2 negative breast cancers develop distant metastases. The metastatic potential of epithelial cells is known to be regulated by tumor–stromal interaction and mediated by epithelial-to-mesenchymal transition. Hormone receptor positive human epidermal growth factor receptor 2 negative tumors were used to estimate markers of epithelial-to-mesenchymal transition, and the luminal breast cancer cell line MCF-7 was used to examine the interactions between integrins and growth factor receptors in causation of epithelial-to-mesenchymal transition. A total of 140 primary tumors were sub-divided into groups enriched for the markers of epithelial-to-mesenchymal transition (snail family transcriptional repressor 2 and integrin β6) versus those with low levels. Within the epithelial-to-mesenchymal transition+ tumors, there was a positive correlation between the transcripts of integrin β6 and growth factor receptors—human epidermal growth factor receptor 2 and epidermal growth factor receptor. In tumors enriched for epithelial-to-mesenchymal transition markers, patients with tumors with the highest quartile of growth factor receptor transcripts had a shorter disease-free survival compared to patients with low growth factor receptor expression by Kaplan–Meier analysis (log rank, p = 0.03). Epithelial-to-mesenchymal transition was induced in MCF-7 cells by treatment with transforming growth factor beta 1 and confirmed by upregulation of SNAI1 and SNAI2 transcripts, increase of vimentin and integrin β6 protein, and repression of E-cadherin. Treatment of these cells with the dual-specificity tyrosine-kinase inhibitor lapatinib led to downregulation of epithelial-to-mesenchymal transition as indicated by lower levels of SNAI1 and SNAI2 transcripts, integrin αvβ6, and matrix metalloproteinase 9 protein. The results suggest that synergistic interactions between growth factor receptors and integrin β6 could mediate epithelial-to-mesenchymal transition and migration in a subset of luminal breast cancers and lapatinib might be effective in disrupting this interaction.

Duodenal gastrointestinal stromal tumor presenting as massive gastrointestinal bleed

Editor Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal (GI) tract, derived from the malignant transformation of the interstitial cells of Cajal or their precursors. GISTs are more common in the 50– 60 years age group and occur commonly in the stomach (60% to 70 %) and small intestine (25 % to 35 %) [1]. Duodenal GISTs are rare, comprising about 12 % to 18 % of small intestine GIST and less than 4 % of all GISTs. They usually present with anemia due to chronic GI blood loss, but duodenal GIST presenting as massive GI bleed with shock is rare. Duodenal GISTs most frequently involve the second portion of the duodenum in close relationship to the ampulla of Vater, followed by the third, fourth, and first portions. The relationship to the ampulla determines surgical treatment strategies. We have treated four patients of GIST presenting with massive upper GI bleed and shock in the last 18 months. Their clinical, laboratory, operative, and histopathology details are outlined in Table 1. All four were between 40–60 years of age. Three out of four presented with hematemesis, and all four were critically ill with hemodynamic instability. Laboratory evaluations revealed severe microcytic hypochromic anemia in all four patients with hemoglobin ranging from 5.6 to 6.8 g/ dL. Coagulation parameters were normal. Gastroduodenoscopy showed submucosal nodule with surface ulceration and active ooze in the second part of the duodenum in three patients and submucosal nodule with active spurt of blood in the third part of the duodenum in one patient (Fig. 1). Hemostasis was achieved with injection of the diluted adrenaline. Contrast-enhanced computed tomography of abdomen confirmed intramural mass in duodenum with distinct fat planes and no lymph node enlargement. All four were operated with limited resection and primary anastomosis. All patients had small size tumors which presented early due to endoenteric growth and had mitosis less than 5 per 50 high-power fields. Their small size, endoenteric growth, complete resection, and lower mitosis were good prognostic signs in our patients. Immunohistochemical analysis revealed a positive CD117 in all four patients, positive CD34 in three patients, and positive smooth muscle actin (SMA) in one. All are alive with no recurrence, with a mean follow up of 10 months ranging from 5 to 18 months. Massive GI bleeding, as in our patients, is unusual in duodenal GIST. In a large study of 156 duodenal GIST patients, 75 cases presented with anemia due to chronic GI loss, but massive GI bleed with hemodynamic instability was only occasional [2]. In the same study, occasional massive GI bleed presented with melena, but none had hematemesis [2]. The typical features of GISTs on duodenoscopy include gross ulceration in the mucosa or an intramural mass with central ulceration. The most specific diagnostic criterion is strong and diffuse positive staining for CD117. GISTs harbor positivity for vimentin in nearly all cases, CD34 in 50 % to 70 %, smooth muscle actin in 30 % to 40 %, and platelet-derived growth factor receptor alpha (PDGFR-α) in about 5 %, while desmin (intermediate filament typical for muscle) and S-100 (a neural cell marker) are usually negative [3]. Poor prognostic parameters of GISTs include extragastric location, size greater than 5 cm, central necrosis, extension into adjacent organs, M. Patil (*) :K. A. Sheth :C. K. Adarsh :H. Devarbhavi Department of Gastroenterology, St. John’s Medical College, Bangalore 560 034, India e-mail: drmalli_arjun@yahoo.co.in

Body image and sexuality in women survivors of breast cancer in India: Qualitative findings

Objectives: With increasing rates of breast cancer survivors, psychosocial issues surrounding cancer survivorship have been gaining prominence. The following article reports on body image and sexuality-related issues in aftermath of the diagnosis and its treatment in the Indian context. Materials and Methods: Research design was mixed method, cross–sectional, and exploratory in nature. Quantitative sample consisted of fifty survivors while the qualitative sample size included 15 out of the 50 total breast cancer survivors who were recruited from hospitals, nongovernmental organization, and through word-of-mouth. Data was collected using quantitative measures, and in-depth interviews were done using semi-structured interview schedule that was developed for the study. Qualitative data were analyzed using descriptive phenomenological approach. Results: In body image, emerging themes were about identity (womanhood, motherhood, and attractiveness), impact of surgery, hair loss, clothes, and uncomfortable situations. In sexuality, barriers were faced due to difficulty in disclosure and themes were about adjustments made by spouses, role of age, and sexual difficulties due to treatment. Conclusions: Findings imply need to address the issues of body image and sexuality as it impacts quality of life of survivors.