Glioma arising in the setting of mismatch repair deficiency-rare or are we missing it?

Germline mutations in mismatch repair (MMR) genes (MLH1, MSH2, MSH6, PMS2) can be mono-allelic or biallelic, resulting in a Lynch syndrome (LS) or constitutional mismatch repair deficiency (CMMRD) syndrome respectively. Glioma arising in the setting of MMR deficiency is uncommon. We describe two pediatric patients with high-grade glioma (HGG) and associated MMR protein deficiency. On histomorphology both cases showed HGG with astrocytic morphology and prominent multinucleated tumor cells. On immunohistochemistry, the first case was negative for IDH1 p.R132H showed loss of ATRX and p53 positivity. The second case was positive for IDH1 p.R132H and p53, but showed retained expression of ATRX. The histomorphology in both cases and additionally IDH mutation with retained ATRX in the second case, prompted us to test for MMR protein deficiency which was carried out by immunohistochemistry (IHC). One case revealed an immunostaining pattern suggestive of CMMRD while the other was suggestive of LS. Both the cases showed good response to surgery and radio-chemotherapy in the follow-up available. Our cases highlight the importance of testing for MMR proteins by simple IHC, in the setting of appropriate clinical scenario, histopathological and immunohistochemical findings. The recognition of these tumors is extremely important to guide further treatment and prompt family screening.

Molecular epidemiology and clinical characteristics of epidermal growth factor receptor mutations in NSCLC: A single-center experience from India.

Background: The genetic profiling of non-small cell lung cancer (NSCLC) has contributed to the discovery of actionable targetable mutations, which have significantly improved outcomes in disease with poor prognosis. Molecular epidemiological data of driver mutations in Indian populations have not been extensively elaborated compared to western and eastern Asian NSCLC populations. This study assessed the prevalence and clinical outcomes of EGFR (epidermal growth factor receptor) mutations among the Indian NSCLC cohort in South India. Patients and Methods: Retrospective analysis of 2,003 NSCLC patients who had undergone EGFR mutational analysis from 2013 to 2020 was performed. Clinical analysis was performed for 141 patients from 2013 to 2017 using Kaplan-Meier and Chi-square methods. Descriptive and survival statistics were performed using IBM SPSS Statistics for Windows, Version 23.0. Armonk, NY: IBM Corp. Results: EGFR-sensitizing mutations were detected in 41.6% (834/2003) in the study cohort with compound mutations detected in 7.55% (63/834) of EGFR-positive cases. A significant relationship with regard to female gender and EGFR mutation status (P <.001) was observed. Exon 18 G719X (8.7%) mutations and exon 20 T790M point mutation (3.1%) were the most frequently isolated uncommon EGFR mutations. In the clinical cohort, EGFR mutations were detected at a significantly higher prevalence in females (P =0.002) and never-smokers (P < 0.001). EGFR mutation demonstrated a significant relationship with regard to brain metastasis (P = 0.011). EGFR mutated individuals had significantly longer median overall survival compared to EGFR wild type (26 months vs. 12 months, P = 0.044). Conclusion: We reports the highest number of EGFR mutation analysis performed from India and mutational analysis indicated a loco-regional variation in India with regard to EGFR mutation frequency and its subtypes.

Recurrent squamous cell carcinoma of the skin treated with immunotherapy.

Squamous cell carcinoma (SCC) has a good prognosis, while metastatic tumors have aggressive behavior. Immunotherapy has become a standard line of treatment in metastatic cancers; pembrolizumab has shown promising results and improved quality of life in recurrent and metastatic cancers. We report a case of recurrent SCC of the skin with extensive disease and a known case of human immunodeficiency virus. He completed standard lines of treatment and currently on immunotherapy. After 3 cycles of immunotherapy plus chemotherapy, he got a complete metabolic response. Our experience showed palliative benefits and increased quality of life.

Excellent long term oncological outcome in a patient with rare glycogen rich clear cell carcinoma of breast following breast conservation surgery.

INTRODUCTION: We present a case report of excellent oncological outcome after 7-year follow up in a female Indian patient with pT2N3aM0 rare GRCC of the breast following breast conservation surgery and appropriate adjuvant treatment. Glycogen rich cell carcinoma (GRCC) is a rare subtype of primary malignant neoplasm of the breast which is not commonly discussed. Only approximately 288 cases have been reported since its first description globally with reports of varying prognosis. Even less (4 patients), which have been reported from India have described only clinic pathological features. This is first case report of patient from India discussing long term oncological outcome of a patient with rare GRCC (pT2N3aMO) of the breast following breast conservation surgery and appropriate adjuvant treatment. A 41-year-old lady presented to us with history of 2 × 2 cm right breast lump for 2 weeks. A BIRAD IV hypo echoic lesion with slightly irregular margins in the upper outer quadrant of the right breast and right axillary lymphadenopathy was reported in mammogram. PET CT showed metabolically active lesion 2.3 × 1.3 cm enhancing nodule with spiculated margins at the same site (SUV-10.8) with metabolically active right axillary metastatic lymphadenopathy (SUV-11) with no distant metastases. Core biopsy indicated Ductal carcinoma. Patient underwent right breast conservation surgery (Wide local excision and oncoplasty with axillary clearance) uneventfully followed by appropriate adjuvant treatment (Chemotherapy, Targeted treatment, Radiotherapy). The final pathological stage was Glycogen rich clear cell carcinoma, pT2N3a M0 with Her2 positive but negative for ER and PR with Ki 67-50 %. The patient had excellent outcome and was alive and cancer free even after 7 years follow up. CONCLUSION: The purpose of reporting this case is to increase the knowledge about this rare subtype of breast cancer which underwent organ preservation. This case report reveals that clinical behavior and oncological outcome of GRCC breast can be unexpected, unusual, varied and even good, contrary to recent 2019 SEER data (Zhou Z, Kinslow CJ, Hibshoosh H, et al. Clinical features, survival and prognostic factors of glycogen-rich clear cell carcinoma (GRCC) of the breast in the US population. J Clin Med. 2019; 8: pii: E246).

Remote Reporting During a Pandemic Using Digital Pathology Solution: Experience from a Tertiary Care Cancer Center.

BACKGROUND: Remote reporting in anatomic pathology is an important advantage of digital pathology that has not been much explored. The COVID-19 pandemic has provided an opportunity to explore this important application of digital pathology system in a tertiary care cancer center to ensure patient care and staff safety. Regulatory guidelines have been described for remote reporting following the pandemic. Herein, we describe our experience of validation of digital pathology workflow for remote reporting to encourage pathologists to utilize this facility which opens door for multiple, multidisciplinary collaborations. OBJECTIVE: To demonstrate the validation and the operational feasibility of remote reporting using a digital pathology system. MATERIALS AND METHODS: Our retrospective validation included whole-slide images (WSIs) of 60 cases of histopathology and 20 cases each of frozen sections and a digital image-based breast algorithm after a washout period of 3 months. Three pathologists with different models of consumer-grade laptops reviewed the cases remotely to assess the diagnostic concordance and operational feasibility of the modified workflow. The slides were digitized on a USFDA-approved Philips UFS 300 scanner at ×40 resolution (0.25 µm/pixel) and viewed on the Image Management System through a web browser. All the essential parameters were reported for each case. After successful validation, 886 cases were reported remotely from March 29, 2020, to June 30, 2020, prospectively. Light microscopy formed the gold standard reference in remote reporting. RESULTS: 100% major diagnostic concordance was observed in the validation of remote reporting in the retrospective and prospective studies using consumer-grade laptops. The deferral rate was 0.34%. 97.6% of histopathology and 100% of frozen sections were signed out within the turnaround time. Network speed and a lack of virtual private network did not significantly affect the study. CONCLUSION: This study of validation and reporting of complete pathology cases remotely, including their operational feasibility during a public health emergency, proves that remote sign-out using a digital pathology system is not inferior to WSIs on medical-grade monitors and light microscopy. Such studies on remote reporting open the door for the use of digital pathology for interinstitutional consultation and collaboration: Its main intended use.

Inflammatory myofibroblastic tumours of the duodenopancreaticobiliary tract.

Inflammatory myofibroblastic tumours (IMTs) are rare benign neoplasms of intermediate malignant potential that are found in the lungs and rarely at extrapulmonary sites common in children and young adults. IMTs tend to be locally invasive and have some amount of metastatic potential as well. We present two cases of IMTs involving the duodenum, pancreas and distal bile duct. The first case presented with extensive involvement of the first three parts of the duodenum and head of the pancreas, while the second presented with a pancreatic and biliary tree involvement. Upon examinations and investigations, these tumours mimicked malignant neoplasms. A Whipple procedure for surgical resection was undertaken in both cases. The histological findings showed fascicles of spindle cells with infiltration of lymphocytes and plasma cells. The inflammatory myofibroblastic tumour was diagnosed based on pathological grounds with immunohistochemistry. Preoperative diagnosis of IMTs is difficult and complete surgical resection is the primary treatment.

Ultrasensitive detection of tumor-specific mutations in saliva of patients with oral cavity squamous cell carcinoma.

BACKGROUND: Oral cavity squamous cell carcinoma (OCSCC) is the most common head and neck malignancy. Although the survival rate of patients with advanced-stage disease remains approximately 20% to 60%, when detected at an early stage, the survival rate approaches 80%, posing a pressing need for a well validated profiling method to assess patients who have a high risk of developing OCSCC. Tumor DNA detection in saliva may provide a robust biomarker platform that overcomes the limitations of current diagnostic tests. However, there is no routine saliva-based screening method for patients with OCSCC. METHODS: The authors designed a custom next-generation sequencing panel with unique molecular identifiers that covers coding regions of 7 frequently mutated genes in OCSCC and applied it on DNA extracted from 121 treatment-naive OCSCC tumors and matched preoperative saliva specimens. RESULTS: By using stringent variant-calling criteria, mutations were detected in 106 tumors, consistent with a predicted detection rate ≥88%. Moreover, mutations identified in primary malignancies were also detected in 93% of saliva samples. To ensure that variants are not errors resulting in false-positive calls, a multistep analytical validation of this approach was performed: 1) re-sequencing of 46 saliva samples confirmed 88% of somatic variants; 2) no functionally relevant mutations were detected in saliva samples from 11 healthy individuals without a history of tobacco or alcohol; and 3) using a panel of 7 synthetic loci across 8 sequencing runs, it was confirmed that the platform developed is reproducible and provides sensitivity on par with droplet digital polymerase chain reaction. CONCLUSIONS: The current data highlight the feasibility of somatic mutation identification in driver genes in saliva collected at the time of OCSCC diagnosis.

Landscape of clinically actionable mutations in breast cancer 'A cohort study'.

Breast cancer (BC) is a heterogeneous disease. Numerous chemotherapeutic agents are available for early stage or advanced/metastatic breast cancer to provide maximum benefit with minimum side effects. However, the clinical outcome of patients with the same clinical and pathological characteristics and treated with similar treatments may show major differences and a vast majority of patients still develop treatment resistance and eventually succumb to disease. It remains an unmet need to identify specific molecular defects, new biomarkers to enable clinicians to adopt individualized treatment for every patient in terms of endocrine, chemotherapy or targeted therapy which will improve clinical outcomes in BC. Our study aimed to identify frequent hotspot mutation profile in BC by targeted deep sequencing in cancer-related genes using Illumina Truseq amplicon/Swift Accel-Amplicon panel and MiSeq technology in an IRB-approved prospective study in a CLIA compliant laboratory. All the cases had pathology review for stage, histological type, hormonal status and Ki-67. Data was processed using Strand NGS™. Mutations identified in the tumor were assessed for 'actionability' i.e. response to therapy and impact on prognosis.

Is a Short-course of Preoperative Denosumab as Effective as Prolonged Therapy for Giant Cell Tumor of Bone?

BACKGROUND: Denosumab is an inhibitor of monoclonal receptor activator of nuclear factor-ĸB ligand, approved to treat giant cell tumors of bone (GCTB). It is commonly used for unresectable tumors and for downstaging the tumor to perform less-morbid procedures. Although denosumab has been used extensively for GCTBs, there are no recommendations regarding the duration of therapy. The risk factors associated with local recurrence (LR) in patients receiving preoperative denosumab for GCTB also are unknown. QUESTIONS/PURPOSES: (1) Is short-course (three doses or fewer) preoperative denosumab treatment as effective as longer course (more than three doses) of treatment in terms of achieving a clinical, radiologic, and histologic response in patients with GCTB? (2) Is there an increased risk of LR after short-course denosumab therapy compared with long-course denosumab therapy; and after controlling for confounding variables, what factors were associated with LR after surgery for GCTB in patients receiving preoperative denosumab? METHODS: A retrospective study was performed using an institutional database of 161 skeletally mature patients with a histologic diagnosis of GCTB who received denosumab between November 2010 and July 2019 to downstage the tumor before surgery. In general, we used denosumab when we thought it would facilitate either resection or curettage (by formation of a sclerotic rim around the osteolytic lesion), when a less-morbid procedure than initially planned might be performed, and in patients with complex presentations like cortical breech and soft tissue extension, pathological fracture, thinning of more than three cortices of the extremity. From 2010 to late 2015, denosumab was administered for approximately 4 to 6 months; starting in late 2015 through 2020, the number of denosumab doses has been reduced. We divided patients into two groups: Those who received three or fewer doses of denosumab (short-course, n = 98) and those who received more than three doses of denosumab (long-course, n = 63). Comparing those in the long-course group with those in the short-course group whose procedures were performed at least 2 years ago, there were no differences in loss to follow-up before 2 years (3% [3 of 98] versus. 3% [2 of 63]). The mean patient age was 30 years (± 6.1) and the mean number of denosumab doses was 4.4 (range 1 to 14). Overall, 77% (37 of 48) of patients taking short-course denosumab and 75% (27 of 36) of patients on long-course denosumab underwent curettage, and the remaining patients with an inadequate bony shell around the tumor or destruction of articular cartilage in both groups underwent tumor resection. With the numbers available, the patients with short- and long-course denosumab were not different in terms of age, sex, MSTS score on presentation, lesion size, lesion location, Campanacci grade, presence of pathological fracture and pulmonary metastasis on presentation, and the type of surgery performed (curettage versus resection). We analyzed the change in the Musculoskeletal Tumor Society score, change in Campanacci grade, radiologic objective tumor response (defined as a partial or complete response, per the modified inverse Choi criteria), and histologic response (defined as reduction of more than 90% of osteoclast-like giant cells or a reduction of more than 50% of mesenchymal spindle-like stromal cells, along with evidence of lamellar or woven bone formation, when compared with the biopsy sample) between the two groups (short- and long-course denosumab). LR rates were compared between the two groups, and after controlling for confounding variables, factors associated with LR in all operated patients were analyzed with a Cox proportional hazards regression analysis. RESULTS: With the numbers available, there was no difference between the short- and long-course denosumab groups in terms of mean percentage improvement in MSTS score (20 [± 18.5] versus 24 [± 12.6]; p = 0.37), radiologic objective tumor response (90% [43 of 48] versus 81% [29 of 36]; p = 0.24) and histologic response (79% [38 of 48] versus 83% [30 of 36]; p = 0.81). With the numbers available, there was no difference between the short- and long-course denosumab groups in terms of Kaplan-Meier survivorship free from LR at 5 years after surgery (73% [95% confidence interval, 68 to 76] versus 64% [95% CI 59 to 68]; log-rank p = 0.50). After controlling for potential confounding variables like age, sex, Campanacci grade and MSTS score on presentation, number of denosumab doses administered before surgery, clinical, radiologic and histologic response to denosumab, and time duration between denosumab therapy and surgery, we found that tumors involving the bones of the hand and the foot (hazard ratio 7.4 [95% CI 2.0 to 27.3]; p = 0.009) and curettage (HR 6.4 [95% CI 2.8 to 23.0]; p = 0.037) were independently associated with a higher risk of LR. CONCLUSIONS: In this preliminary, single-center study, we found that a short-course of preoperative denosumab (three or fewer doses) was associated with no differences in clinical scores, histological and radiological response, or LR-free survivorship, compared with longer-course of denosumab (more than three doses). Fewer preoperative doses can reduce the complications and costs associated with more-prolonged therapy. Denosumab must be used cautiously before curettage for GCTB, and only if the benefit of joint salvage outweighs the possibility of LR. However, given the small number of patients, potentially clinically important differences might have been missed, and so our findings need to be confirmed by larger, multicenter, prospective trials. LEVEL OF EVIDENCE: Level III, therapeutic study.

Incidence of Micrometastasis and Isolated Tumour Cells in Clinicopathologically Node-Negative Head and Neck Squamous Cell Carcinoma.

INTRODUCTION: Occult neck node metastasis in head and neck squamous cell carcinoma (HNSCC) in the form of micrometastasis and isolated tumour cell (ITC) often goes unnoticed in the routine pathological examination. This limitation can be overcome by using serial sectioning and immunohistochemistry for detection of micrometastasis and ITC in clinically and pathologically node-negative neck. The primary objective was to determine the incidence of micrometastasis and ITC in the selective neck dissection specimen, whereas to determine the levels of lymph nodes involved, depending upon the site of primary tumour, was the secondary objective. MATERIALS AND METHODS: Lymph nodes from selective neck dissection specimen were subjected to serial sectioning and immunohistochemistry with pan-cytokeratin marker. Incidence of micrometastasis and ITC, site and stage of primary tumours and level of lymph nodes involved were determined. RESULTS: In total, 8.8% patients in the study got upstaged after serial sectioning and immunohistochemistry. Tongue and lower alveolar primaries showed the presence of micrometastasis and ITC in neck nodes. All the primary tumours were of pT1 stage. Level IB and II lymph nodes were primarily involved. CONCLUSION: Micrometastasis and isolated tumour cells are found in approximately 9% of cases of early-stage oral cavity squamous cell carcinoma. The predictive factors and clinical significance are still unknown. More prospective trials are required to solve this evolving aspect of HNSCC.

Nascent verrucous hyperplasia - A transition to cutaneous squamous cell carcinoma.

Cutaneous squamous cell carcinoma is a common global cancer with Ultraviolet light recognized as the most significant risk factor. The other definite or plausible risk factors include immunosuppression, infection with oncogenic viruses, exposure to toxins, chemicals, chronic inflammatory skin disease and a high body mass index. This case highlights the rarity of the pathology in terms of size, the subtle transition of verrucous hyperplasia to cutaneous squamous cell carcinoma over a period of time and the fallibility of the frozen section report in deciding the optimum resection margins. The initial innocuous presentation represented a diagnostic challenge as it can be mistaken for other benign entities. A correlation between the history, clinical presentation, tumor biology and the histopathological characteristics helped us to unlock the jigsaw puzzle of approaching a rare condition with a modification in the surgical approach.

Case of Solitary Peritoneal Metastasis From Atypical Bronchopulmonary Carcinoid on 18F-FDG PET/CT.

Atypical lung carcinoids frequently metastasize to mediastinal nodes, liver, bone, lungs, and brain and rarely to ovaries, pancreas, subcutaneous, and skin. Solitary peritoneal metastasis is extremely rare and unreported previously. We present a case of a 36-year-old woman with cough, hemoptysis, dyspnea with detection of lung mass, and incidental detection of pelvic mass. Fine needle aspiration cytology failed to reveal the true nature of both the lesions. FDG PET/CT showed intensely avid mediastinal nodes, left lung, and rectouterine masses. Core biopsy from lung mass revealed atypical carcinoid without appreciable uptake on Ga-DOTANOC scan. Subsequent postchemotherapy CT scan shows significant regression in lung and peritoneal masses.

Can low grade PMP be divided into prognostically distinct subgroups based on histological features? A retrospective study and the importance of using the appropriate classification.

INTRODUCTION: The pathological classification of PMP of appendiceal origin has prognostic and treatment implications. Our goals were to • Classify low grade mucinous carcinoma peritonei (LGMCP) into prognostically distinct subgroups based on histological features. • Compare the reproducibility of the WHO and the PSOGI classifications for both PMP and the appendiceal primary tumor. PATIENTS AND METHODS: A retrospective analysis of patients undergoing CRS and HIPEC or debulking surgery was done. All the tumors were re-classified according to the PSOGI classification. LGMCP was further classified into three histological subgroups and the impact on survival was evaluated. RESULTS: From Jun 2011 to June 2016, 101 patients underwent CRS with HIPEC (n = 89) or debulking surgery (n=12). The median PCI was 28 (3-39) and 74.1% patients had CC-0/1 resections. Of the 76.2% patients who had LGMCP, 4 patients (5.1%) were classified as group 1, 54 (70.1%) as group 2 and 19 patients (24.6%) as group 3. At a median follow up of 21 months, the disease free survival was not reached, 30 months and 14 months for groups 1, 2 and 3 respectively (p = 0.09). There was no difference in overall survival. Using the WHO classification, there was a discordance in the grade of the primary tumor and the peritoneal lesions in 19.8% and conflicting terminology was used in 62% of patients. CONCLUSIONS: The subgroups of LGMCP described here are prognostically different though this needs further prospective evaluation in larger series. The PSOGI classification is more uniformly reproducible and should be preferred to the WHO classification.

Outcomes of cytoreductive surgery and HIPEC for pseudomyxoma peritonei of appendiceal origin from two Indian centers: A preliminary five-year experience.

PURPOSE: To evaluate the short-term outcomes of patients of pseudomyxoma peritonei (PMP) of appendiceal origin treated with cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) at two tertiary Indian centers. METHODS: Data was prospectively collected from January 2011 to January 2016. Palliative procedures were excluded. HIPEC was performed by the coliseum technique using either a mitomycin or oxaliplatin-based regimen. RESULTS: 77 procedures were performed on 71 patients. The average time interval between diagnosis and CRS was 15.3 months. Of the tumors, 22.1% were high grade, 77.9% low grade and 24.6% intermediate grade. The median peritoneal carcinomatosis index (PCI) was 26 (<25 in 70.1% and >30 in 38.9%). Completeness of cytoreduction score (CCS)-1 was achieved in 75.3% (CC-0 in 42.9%). The mean number of bowel anastomoses was 1.1 and the mean number of organs resected per patient was 3.3. Of the 77 patients, 71% had resection of 3 or more organs and 50.6% had resection of 4 or more organs. Grade 3-4 complications occurred in 42.9% of the patients and the perioperative mortality was 5.2%. The projected 5-year overall survival (OS) was 62.3% and the 3-year disease-free survival (DFS) was 71% at a median follow up of 13 months. CONCLUSION: CRS and HIPEC can be used to treat PMP with an acceptable morbidity and mortality in Indian patients. Lack of early referrals leads to a large portion of patients presenting with extensive disease and an inferior survival which should improve with increasing awareness about the procedure and its results.

Pathology of Mucinous Appendiceal Tumors and Pseudomyxoma Peritonei.

Neoplasms of the appendix are rare, but because of their unusual presentation and unpredictable biologic behavior, it is important to diagnose them correctly. Mucinous tumors account for 58 % of malignant tumors of appendix in SEER database and the remaining are carcinoids. The mucinous appendiceal tumors have a potential to spread to the peritoneum and viscera in the form of gelatinous material with or without neoplastic cells resulting in Pseudomyxoma peritonei. (PMP) PMP is a clinical entity that has a unique biological behavior and can arise from seemingly benign tumors to frankly malignant ones. Several classifications exist for PMP of which Ronnet's classification has been the most popular. In 2010, the WHO proposed a 2 tier classification that classified PMP as either low grade or high grade based on the presence of mucin, cytological and architectural features. According to this classification when the underlying cause for PMP is an appendiceal tumor it is always a mucinous adenocarcinoma rather than a mucocoele or adenoma and these terms should no longer be used. This system of classification helps in predicting the behavior of the tumor and proper treatment strategies. The understanding of the pathogenesis of the disease has also improved with identification of newer biomarkers and molecular genetic alterations. IHC markers CK 20, CDX2 and MUC2 are found to be positive in these tumors in addition to KRAS mutation and loss of heterozygosity in some gene loci. Proper histopathologic classification and predicting the tumor behavior requires a close interaction between the pathologist and the surgeon. The use of the combined modality treatment of cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) has led to a 5-year survival ranging from 62.5 % to 100 % for low grade, and 0 %-65 % for high grade disease. This article focuses on the etiopathogenesis, clinical behavior, diagnosis and classification of mucinous tumors of the appendix and pseudomyxoma peritonei.