Amal C Kataki

Imbalance in leptin-adiponectin levels and leptin receptor expression as chief contributors to triple negative breast cancer progression in Northeast India

BACKGROUND Triple-Negative breast cancer (TNBC), accounts for a large percentage of breast cancer cases in India including Northeast India. TNBC has an unclear molecular aetiology and hence limited targeted therapies. Human breast is comprised of glandular, ductal, connective, and adipose tissues. Adipose tissue is composed of adipocytes. The adipocytes apart from being energy storage depots, are also active sources of adipocytokines and/or adipokines. The role of adipokines in breast cancer including TNBC has been sporadically documented. Two adipokines in particular, leptin and adiponectin, have come to be recognized for their influence on breast cancer risk and tumour biology. Therefore, the aim of this study was to understand the association of differential expression of critical adipokines and associated cellular mechanism in the susceptibility and severity of TNBC in northeast Indian population. MATERIALS AND METHODS We collected 68 TNBC and 63 controls cases and examined for serum leptin and adiponectin levels using enzyme linked immunosorbent assay (ELISA). Leptin Receptor (Ob-R) mRNA expression was determined by real-time polymerase chain reaction (RT-PCR) assay. Differential Ob-R mRNA expression and correlation with cancer stem cell (CSC) markers was evaluated, and correlated with severity. RESULTS The serum leptin levels were significantly associated with TNBC severity, while the adiponectin levels were comparative. The serum leptin levels correlated inversely with the adiponetin levels. Serum leptin levels were unaffected with difference in parity. The difference in leptin levels in pre and post menopausal cases were found to be statistically non-significant. Higher leptin levels were also found to be associated obesity, mortality and recurrence. Obesity was found to be a factor for TNBC pathogenesis and severity. Increased Ob-R mRNA expression was associated with TNBC, significantly with TNBC severity, and was significantly higher in obese patients with higher grade TNBC cases. The Ob-R gene mRNA expression was significantly higher in the obese TNBC cases showing recurrence or mortality. The higher Ob-R gene mRNA expression correlated significantly with higher serum leptin levels and lower serum adiponectin levels in TNBC cases. The Ob-R mRNA expression with associated with modulation of CSC oct4 and nanog. CONCLUSIONS In conclusion, the present study is first of its kind on TNBC from northeast India, indicates that adipocytokines does play a role in TNBC pathogenesis. Thus, the understanding of molecular mechanisms of both leptin and adiponectin and their interplay in TNBC offer the prospects for new therapeutic approaches targeting similar signalling pathways.

Inflammatory myofibroblastic tumor of the upper alveolus: A rare entity presenting as a jaw swelling

Inflammatory myofibroblastic tumor (IMT) is a rare tumor of borderline nature that can clinically present as a malignant neoplasm. It commonly occurs in the lungs, and a very few oral IMTs have been reported in the literature. IMT consists of inflammatory cells and myofibroblastic spindle cells. The diagnosis of IMT requires histopathological examination with immunohistochemical staining to look for the expression of smooth-muscle actin for confirmation of the diagnosis. The objective of this paper is to report an IMT on the upper alveolus with clinic-a pathological similarity with a malignant lesion and its management. Though oral IMTs are rare, it should be considered in the differential diagnosis of tumors of the upper jaw. Complete surgical excision of alveolar IMT is the treatment of choice because of its unpredictable clinical behavior. The patients with oral IMTs require periodic post-surgical follow-up for recurrence.

Cutaneous metastasis from squamous carcinoma of the base of tongue

Context: Cutaneous metastasis from head and neck cancer is uncommon and it is seen from laryngeal cancer. Cutaneous metastasis from the base of tongue is relatively rare. Case Report: A 55-year-old male, who was a treated case of squamous carcinoma of the base of tongue presented with metastatic nodule on the skin of face and thigh. But, there was complete resolution of the tumor at the primary site. In the present case, clinically obvious cutaneous nodules with metastasis appeared soon after the completion of treatment with concurrent chemo-radiotherapy. The metastasis to the skin of face clinically appeared like an inflammatory lesion. Fine needle aspiration cytology confirmed the diagnosis of metastasis to skin at both the sites. Conclusion: Our case has highlighted that there could be associated occult skin metastasis at the time of diagnosis in squamous carcinoma of the base of tongue.

Papillary thyroid carcinoma presenting with mandibular metastasis: An unusual presentation

Mandibular metastasis from papillary carcinoma of the thyroid is extremely rare. We report here a case of metastatic swelling on the mandible due to papillary carcinoma of the thyroid. The patient presented with jaw swelling and the thyroid lesion was an incidental finding on clinical examination. Computed tomogram scan revealed the presence of a contrast enhanced lesion in the thyroid and lytic expansile lesion in the body of the mandible. The diagnosis of papillary thyroid carcinoma (PTC) with mandibular metastasis was made after cytological examination of both the lesions. The patient was treated with surgery followed by radioiodine ablation. In conclusion, metastatic tumor to the jaw from a PTC is an extremely rare phenomenon and in the differential diagnosis of a metastatic jaw swelling small primary tumors of the thyroid should be excluded.

Abstract A09: Role of adipocytokines in obesity associated Triple Negative Breast Cancer

Background: Triple-negative breast cancer (TNBC), accounts for a large percentage of breast cancer cases in India including Northeast India. TNBC has an unclear molecular aetiology and hence limited targeted therapies. Human breast is comprised of glandular, ductal, connective, and adipose tissues. Adipose tissue is composed of adipocytes. The adipocytes apart from being energy storage depots are also active sources of adipocytokines and/or adipokines. The role of adipokines in breast cancer including TNBC has been sporadically documented. Two adipokines in particular, leptin and adiponectin, have come to be recognized for their influence on breast cancer risk and tumor biology. Therefore, the aim of this study was to understand the association of differential expression of critical adipokines and associated cellular mechanism in the susceptibility and severity of TNBC in northeast Indian population. Materials and Methods: We collected 68 TNBC and 63 controls cases and examined for serum leptin and adiponectin levels using enzyme linked immunosorbent assay (ELISA). Leptin Receptor (Ob-R) mRNA expression was determined by real-time polymerase chain reaction (RT-PCR) assay. Differential Ob-R mRNA expression and correlation with cancer stem cell (CSC) markers oct4 and nanog was evaluated, and correlated with severity. Results: The serum leptin levels were significantly associated with TNBC severity, while the adiponectin levels were comparative. The serum leptin levels correlated inversely with the adiponetin levels. Serum leptin levels were unaffected with difference in parity. The difference in leptin levels in pre and post menopausal cases were found to be statistically non-significant. Higher leptin levels were also found to be associated obesity, mortality and recurrence. Obesity was found to be a factor for TNBC pathogenesis and severity. Increased Ob-R mRNA expression was associated with TNBC, significantly with TNBC severity, and was significantly higher in obese patients with higher grade TNBC cases. The Ob-R gene mRNA expression was significantly higher in the obese TNBC cases showing recurrence or mortality. The higher Ob-R gene mRNA expression correlated significantly with higher serum leptin levels and lower serum adiponectin levels in TNBC cases. The Ob-R mRNA expression with associated with modulation of CSC. Conclusions: In conclusion, the present study is first of its kind on TNBC from northeast India, indicates that adipocytokines does play a role in TNBC pathogenesis. Thus, the understanding of molecular mechanisms of both leptin and adiponectin and their interplay in TNBC offer the prospects for new therapeutic approaches targeting similar signaling pathways. Note: This abstract was not presented at the conference. Citation Format: Rizwana Sultana, Amal Ch Kataki, Bibhuti Bhusan Barthakur, Sujoy Bose. Role of adipocytokines in obesity associated Triple Negative Breast Cancer [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr A09.

Abstract 2681: Oral micro biome enhances stemness in oral cancer cells by activating Toll like receptor signaling

Background: The resistance and progression of cancers after chemotherapy to invasive and metastatic stages accounts for the overwhelming majority of cancer deaths. Recent studies suggest, microbiomes can induce a cascade of host events to either support or inhibit tumor growth. Specially, in oral cancer, chemotherapy treatment may alter the oral microbial flora, which may favor or inhibit tumor growth. Hence, it is importantl to develop novel experimental approaches to study the role of oral microbial flora in oral cancer stemness (self-renewal and undifferntiated state of cancer stem cells). Importantly, patients in developing area, including Assam, where KaviKrishna laboratory is located, may have distinct oral microbial flora that could favor oral cancer growth. Hence, it is important to include patients from developing countries for such studies. Our previous research showed that chemotherapy ehances stemenss in many cancer cell types, including oral squamous cell carcinoma cell line SSC-25. The stemness switch is characterized by enhanced expression of stemness associated genes including Nanog, Lin28A/B, Oct-4, MYC, HIF-2alpha and inflammation associated genes including Toll like receptor (TLR) 2/4. Here we investigated the role of oral microbiomes in the TLR mediated stemness switch of oral cancer cells. Methods: SCC-25 oral cancer cell line was treated with bacterial product lipopoly saccharide (LPS), and the stemness switch evaluated by isolation of ABCG2+ cells and expression of stemness associated genes by these cells. Capacity of interaction of tumor stromal cells with mesenchymal stem cells was also evaluated. Additionally, we obtained sputum from oral cancer subjects undergoing chemotherapy. The patients were from the Kamrup district of Assam, where KaviKrishna laboratory is located. The sputum was processed and then added to the culture medium of SCC-25 cells. These post-sputum treated SCC-25 cells were subjected to phenotypic stemness switch analysis. Results: We found, LPS and sputum treatment led to the enhanced stemness of ABCG2+ cells, including the high expression of TLR2/4, MYC, Nanog, Sox-2, and HIF-2alpha. Importantly, sputum derived from oral cancer subjects under remission showed inhibitory activity on ABCG2+ cell self-renewal. In contrast, sputum obtained from oral cancer subjects with relapse showed enhanced stemness of ABCG2+ cells, and also increased tumorigenic potential. The post-sputum treated ABCG2+ cells exhibited high expression of TLR2/4 and associated increase of HIF-2alpha and MYC transcriptional activity. The sputum treated with broad spectrum antibiotic ciprofloxacin did not enhance the stemness and TLR2/4 signaling of SCC-25 cells. Conclusion: These results indicate that oral microbiomes may differentially influence the stemness of oral cancer cells. We also conclude that live bacteria present in the sputum may be required to enhance stemness in a TLR2/4 dependent manner. Note: This abstract was not presented at the meeting. Citation Format: Joyeeta Talukdar, Rashmi Bhuyan, Bidisha Pal, Sorra Sandhya, Hong Li, Seema Bhuyan, Sukanya Garhyan, Debabrat Baishya, Anupam Sarma, Jyotirmoy Phukan, Amal Kataki, Bikul Das. Oral micro biome enhances stemness in oral cancer cells by activating Toll like receptor signaling [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 2681. doi:10.1158/1538-7445.AM2017-2681

Abstract 4324: Undenatured whey protein isolate exhibit chemopreventive activity against HPV-16 induced carcinogenesis of CD271+ oral mucosa stem cells

Oral cancer, and associated HPV-16 infection, and tobacco smoking is a major public health concern in the Kamrup district of India. To facilitate research on oral cancer chemoprevention, we have set up a collaborative research team between Forsyth Institute, an oral medicine specialized center affiliated to Harvard School of Dental Medicine, and KaviKrishna Laboratory, a non-for profit private laboratory set up located in Guwahati, the major city of Kamrup district, India. To study the cellular and molecular mechanism of HPV-16 mediated carcinogenesis, we have developed an in vitro model of HPV-16 mediated carcinogenesis of oral squamous stem cells. In this model (Bhuyan et al. The potential role of oral mucosa stem cell altruistic behavior as the initiating event of malignant transformation. AACR abstract Control # 16-A-6618-AACR), the treatment of oral mucosa cells derived from healthy volunteer with HPV-16 derived E6 protein led to expansion of a p53 deficient CD271+ expressing oral mucosa stem cells (OMSC). Noted that CD271 cell surface marker was recently identified as a putative marker for OMSCs. Using this in vitro carcinogenesis model, we evaluated the potential chemopreventive role of dietary whey protein. The E6 treated p53 deficient CD271+ cells were treated with DMEM/F12 media containing 0.2% of Immunocal, an undenatured whey protein (Tsai WY et al. Nutr Cancer 2000. PMID: 11525598). The p53 status as well as in vitro self-renewal activity of the CD271+ cells were examined by ELISA, transcriptional activity assay, and methylcellulose based clonogenic assay. We also fed Immunocal 20 gm/daily for 6 months to 5 individual with oral leukoplakia lesion with HPV-16 positivity. The CD271+ cells obtained from the leukoplakia lesion of clinical subjects were subjected to p53 status, and clonogenic assay. The data was compared with CD271+ cells obtained from leukoplakia lesion of subjects taking regular diet without whey protein supplements. We found that addition of Immunocal whey protein led to 8-fold decrease in the expansion of CD271+ cells following E6 treatment in the in vitro model of OMSC culture. Importantly, Immunocal treatment prevented the suppression of p53 in OMSCs. In the preliminary clinical study, the dietary intake of Immunocal led to complete loss of leukoplakia lesion in the 4/5 individual. In contrast, the control subjects exhibited the presence of CD271+ cells in the leukoplakia lesion having low p53 status, and high clonogenic activity. We are incorporating a larger number of subjects to study the potential chemopreventive activity of whey protein, using the locally available affordable whey protein extracts. Conclusion: This study indicate that whey protein extract, which is available in the local villages of Kamrup district could serve as a chemopreventive agent against oral cancer. Citation Format: Sora Sandhya, Rika Tsuchida, Sukanya Gayan, Rashmi Bhuyan, Joyeeta Talukdar, Bidisha Pal, Seema Bhuyan, Jugal Ch Das, Amal Ch Kataki, Debabrata Baishya, Bikul Das. Undenatured whey protein isolate exhibit chemopreventive activity against HPV-16 induced carcinogenesis of CD271+ oral mucosa stem cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4324.

Abstract 251: Stem cell altruism may serve as a novel drug resistance mechanism in oral cancer

Background: The mechanism of oral squamous cell carcinoma resistance to platinum-based chemotherapy is not clearly known. Previous studies indicated that glutathione (GSH), a cellular antioxidant may detoxify cisplatinum (CDDP), a commonly used chemotherapy agent in oral cancer. Our previous research in human embryonic stem cells (hESCs) indicated the altruistic behavior of ABCG2+ hESCs that secrete high level of GSH to protect other hESCs exposed to oxidative stress (Das B et al. Stem Cells, 2012). Here we investigated if CDDP exposure lead to altruistic stem cell reprogramming of ABCG2+ oral squamous cell carcinoma cells and subsequent GSH-mediated resistance against CDDP. Methods: Two oral squamous cell cancer cell lines SCC-25 and SCC-15 were treated with 3-10 uM CDDP for three-days, and then subjected to flow cytometry and immunomagnetic sorting based evaluation of ABCG2+ cells. The conditioned media (CM) obtained from ABCG2+ cells were examined for GSH content. The CM treated cancer cell lines were examined for resistance against CDDP toxicity. Next, the post-CDDP treated ABCG2+ cells were examined for enhanced stemness phenotype that corresponds to altruistic stem cell phenotype (Das B et al, Stem Cells 2012). Results: We found that CDDP treatment increases the ABCG2+ self-renewal capacity of SCC-25 and SCC-15 cells as measured by serial transplantation assay. The CM of the post-CDDP treated cells exhibited high level of GSH. When the SCC-15 and SCC-25 cells were treated with CM plus CDDP, the cancer cells exhibited 10-15-fold increase in resistance against CDDP toxicity. Next, the post-CDDP treated SCC-25 and SCC-15 cells exhibited enhanced stemness reprogramming phenotype characterized by very high HIF-2alpha, Sox-2 and Nanog transcriptional activity. Furthemore, we found increased expression of EMT (epithelial mesenchymal transition) marker expression including Snail, Twist and vimentin as evaluated by flow cytometry. siRNA HIF-2alpha treatment led to marked loss in the in vivo self-renewal capacity of ABCG2+ SCC-25 and SCC-15 cells. We then noted that post-CDDP ABCG2+ cells exhibited reversible state, as after two weeks of culture, most of the cells either differentiated or underwent apoptosis. Conclusions: These results indicate that oral cancer cells exhibit altruistic defense mechanism to resist the toxicity of CDDP. The altruistic defense mechanism involved high secretion of GSH. Thus, we suggest that similar to bacterial altruism as a mechanism of drug resistance, stem cell altruism may also serve as a novel mechanism of drug resistance in cancer. Citation Format: Bidisha Pal, Reza Bayat-Mokhtari, Hong Li, Rashmi Bhuyan, Joyeeta Talukdar, Sora Sandhya, Anupam Sarma, Wael Tasabehji, Seema Bhuyan, Sukanya Gayan, Amal Ch Kataki, Debabrata Baishya, Herman Yeger, Dean W. Felsher, Bikul Das. Stem cell altruism may serve as a novel drug resistance mechanism in oral cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 251.

Epithelial myoepithelial carcinoma of the lower alveolus

Epithelial-myoepithelial carcinoma (EMC) is a biphasic tumor and classified as a low-grade tumor of the salivary glands. EMC commonly occurs in the parotids with a reported incidence of around 1% out of all neoplasms of the salivary glands and it is a rare tumor of the minor salivary glands in the oral cavity, especially on the lower alveolus. A 48-year-old male patient presented with a rapidly progressive ulcer-infiltrative growth on the lower alveolus with involvement of the overlying cheek skin in a short span of period. We describe here a case of locally advanced EMC of the lower alveolus, its diagnosis by histopathological examination and immunohistochemical staining, and management by surgery and local external beam radiotherapy (EBRT). In our limited experience in the treatment of EMC on the lower alveolus, wide local excision of the tumor, followed by EBRT had resulted in a good therapeutic outcome at the 3 years posttreatment follow-up. In conclusion, EMC of the lower alveolar mucosa is extremely rare, and patients should also be advised close follow-up for the detection of loco-regional recurrences after treatment.

Basaloid squamous carcinoma of skin associated with xeroderma pigmentosum in an 8-year-old child: A rare entity

Xeroderma pigmentosum (XP) is a rare autosomal recessive genodermatosis associated with hypersensitivity to ultraviolet (UV) light, due to defects in deoxyribonucleic acid (DNA) repair. Basaloid squamous cell carcinoma is a rare aggressive variant of squamous cell carcinoma. Patients with XP are at increased risk of developing cutaneous malignancy and are commonly associated with squamous carcinoma. We report an extremely rare case of 8-year-old child with XP along with basaloidsquamous carcinoma of skin; and review of literature related to it.