HPV51-associated Leiomyosarcoma: A Novel Class of TP53/RB1-Wildtype Tumor With Predilection for the Female Lower Reproductive Tract.

Inactivating mutations in tumor suppressor genes TP53 and RB1 are considered central drivers in leiomyosarcomas (LMSs). In high-risk human papillomavirus (HPV)-related tumors, a similar functional outcome is achieved through oncoproteins E6 and E7, which inactivate the p53 and RB1 proteins, respectively. Here, we hypothesized that HPV infection could provide an alternative mechanism for tumorigenesis in a subset of TP53/RB1-wildtype LMS. We evaluated tumor samples from 2585 consecutive unique patients carrying a diagnosis of gynecologic or soft tissue LMS. Tumor DNA and available RNA were analyzed by hybrid-capture-based next-generation sequencing/comprehensive genomic profiling of 406 genes and transcripts (FoundationOneHeme). Of the initial 2585 cases, we excluded 16 based on the presence of molecular alterations that are considered defining for sarcomas other than LMS. In the remaining 2569 cases, we searched for LMS that were TP53/RB1-wildtype (n=486 of 2569; 18.9%). We also searched LMS tumors for HPV sequences that we then classified into genotypes by de novo assembly of nonhuman sequencing reads followed by alignment to the RefSeq database. Among TP53/RB1-wildtype LMS, we identified 18 unique cases harboring HPV sequences. Surprisingly, most (n=11) were HPV51-positive, and these 11 represented all HPV51-positive tumors in our entire LMS database (n=11 of 2569; 0.4%). The absence of genomic alterations in TP53 or RB1 in HPV51-positive LMS represented a marked difference from HPV51-negative LMS (n=2558; 0% vs. 72% [P<0.00001], 0% vs. 53% [P=0.0002]). In addition, compared with HPV51-negative LMS, HPV51-positive LMS were significantly enriched for genomic alterations in ATRX (55% vs. 24%, P=0.027) and TSC1 (18% vs. 0.6%, P=0.0047). All HPV51-positive LMS were in women; median age was 54 years at surgery (range: 23 to 74 y). All known primary sites were from the gynecologic tract or adjacent anogenital area, including 5 cases of vaginal primary site. Histology was heterogeneous, with evaluable cases showing predominant epithelioid (n=5) and spindle (n=5) morphology. In situ hybridization confirmed the presence of high-risk HPV E6/E7 mRNA in tumor cells in three of three evaluable cases harboring HPV51 genomic sequences. Overall, in our pan-LMS analysis, HPV reads were identified in a subset of TP53/RB1-wildtype LMS. For all HPV51-associated LMS, the striking absence of any detectable TP53 or RB1 mutations and predilection for the female lower reproductive tract supports our hypothesis that high-risk HPV can be an alternative tumorigenic mechanism in this distinct class of LMS.

Multi-institutional study of fine-needle aspiration for thyroid lymphoma.

INTRODUCTION: Primary thyroid lymphoma is a rare neoplasm accounting for 1% to 5% of thyroid malignancies. We study the efficacy of fine-needle aspiration (FNA) in diagnosing thyroid lymphoma. METHODS: Pathology databases from our three institutions were searched for thyroid FNA biopsies having a diagnosis of lymphoma or atypical lymphoproliferative cells, or a corresponding tissue diagnosis of thyroid lymphoma having a prior FNA biopsy. RESULTS: Sixty-eight cases were retrieved from 64 patients; 67 cases with histologic confirmation. Forty-six specimens were from women (68%), ages 21 to 87 years (mean = 60). Forty-seven aspirates were diagnosed as lymphoma (n = 29) or suspicious (n = 18) for lymphoma (sensitivity = 73%), 11 atypical, 7 benign, 2 unsatisfactory, and 1 suspicious for carcinoma. Follow-up surgical diagnoses included diffuse large B-cell lymphoma (n = 43), classical Hodgkin lymphoma (5), chronic lymphocytic leukemia (5), and other cases (11). Only 12 of 64 patients (13 specimens) had a known diagnosis of lymphoma prior to FNA. Light chain restriction was detected in 34 specimens (by flow cytometry [FCM] in 32 cases or polymerase chain reaction, in 2 cases). FCM was polyclonal (n = 7) or inconclusive (2) with 25 cases not having FCM performed or not having enough viable cells for evaluation. Four cases showed lymphocytic thyroiditis on surgical follow-up with 2 of these cases having a small monoclonal lymphoid population detected by FCM. CONCLUSIONS: Diffuse large B-cell lymphoma was the most common lymphoma in this series (63%). The sensitivity of FNA with the optional use of FCM was 71% with a specificity of 99%.

Aptima HR-HPV testing from Diff-Quick-stained fine-needle aspiration smears of oropharyngeal squamous cell carcinoma.

INTRODUCTION: Human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma (SCC) is a biologically unique form of carcinoma that is important to identify for prognosis and treatment. The objective of this study was to evaluate the performance of the Aptima HPV assay using Diff-Quick (DQ) stained smears from fine-needle aspiration (FNA) of HPV-related oropharyngeal SCC. MATERIALS AND METHODS: Patients with a diagnosis of head and neck SCC who also had FNA sample demonstrating metastatic disease were identified. Using a mounting media-based cell transfer technique, approximately 200 tumor cells were selected and harvested from DQ-stained aspirate smeared slides. The selected cells were tested for high risk HPV using the Aptima HPV assay, an in vitro nucleic acid amplification test for the qualitative detection of E6/E7 viral messenger RNA from high-risk types of HPV. These results were compared with the p16 immunohistochemical staining of the corresponding surgical pathology specimens. RESULTS: Twenty-eight of 32 (87.5%) FNAs of p16-positive oropharyngeal SCC were positive for high-risk HPV by the Aptima assay and 18 of 18 (100%) FNAs of p16-negative SCC were negative for high-risk HPV by the Aptima assay. CONCLUSIONS: DQ-stained FNA smears can be used by the Aptima HPV assay to accurately detect high-risk HPVs in oropharyngeal SCCs with a sensitivity of 87.5% and a specificity of 100%. This provides an alternative to p16 immunohistochemical staining of FNA cell block material, which may not be available on all specimens.

Correlation of liver and pancreas endoscopic ultrasonography-guided fine-needle aspiration biopsy in patients with a primary pancreatic lesion.

BACKGROUND: Endoscopic ultrasonography (EUS) is commonly used in the evaluation of pancreas masses, and when a liver lesion is visualized, it can undergo a fine-needle aspiration (FNA). This can provide diagnostic and staging information. The purpose of the study was to correlate the findings of patients who underwent EUS FNA biopsy of a pancreas lesion and a liver lesion during the same procedure. MATERIALS AND METHODS: The pathology database at Washington University Medical Center was searched for EUS FNA biopsy cases where biopsy of both the pancreas and liver were performed over a consecutive 10-year period (2003-2013). All pathology reports were reviewed, and clinical information and diagnostic results were recorded. RESULTS: A total of 102 cases were identified. For pancreas cases, 79.4% were malignant and for liver cases, 58.8% were malignant. In pancreas lesions categorized as suspicious for malignancy (9%), the liver biopsy provided a diagnosis of malignancy in 67% of cases. A malignant pancreatic cohort demonstrated a 62.9% liver malignancy. A malignant liver cohort corresponded to a malignant pancreas diagnosis in 86.6% of cases and a suspicious-malignant group of 98.3%. CONCLUSIONS: The 102 cases with concomitant EUS FNA biopsy of the pancreas and liver demonstrated the ability to provide a diagnosis of pancreas malignancy and correlate regional metastatic malignancy in the liver. In patients with a pancreas mass and in the appropriate clinical setting, a liver EUS FNA biopsy has the ability to provide a diagnosis of malignancy and demonstrate a high positive predictive value of malignancy in the pancreas (98.3%).

Squamous cells in effusions: A study of 24 cases.

INTRODUCTION: Malignant squamous cells in serous effusions are rare. We present our experience with squamous cell carcinoma (SqCC) and benign squamous cells in effusions. MATERIALS AND METHODS: Specimens were retrieved from our database using search codes as "squamous" within the final diagnosis under various serous effusions. RESULTS: Twenty-nine specimens were recovered, and 5 of those were excluded. Of the 24 specimens, 3 were duplicates; therefore, only the first specimen from each was included for a final tally of 21 specimens. Specimens were from pleural fluid (n = 16, 76%), pericardial fluid (n = 2), pelvic fluid (n = 2), and peritoneal fluid (n = 1). Nineteen were SqCC (primary sites: 7 lung, 2 uterine cervix, 2 larynx, 2 anus, 2 esophagus, 1 tongue, 1 mandible, 1 skin, 1 vulva), and 2 patients had benign squamous cells only (1 from a ruptured esophageal adenocarcinoma and 1 from a nonmalignant esophageal rupture). In SqCC cases, a round-oval cell with dense cytoplasm was the predominant cell type (n = 12) followed by undifferentiated cells (n = 4), polygonal-type cells (n = 2), and fiber-type cells (n = 1). Of the SqCC specimens, 12 (63%) showed varying degrees of keratinization. Other features in SqCC cases included refractile rings (89%), keratin pearls (53%), and vacuolated cytoplasm (42%). Herxheimer spirals were absent. Two benign cases showed polygonal cell morphology only. All patients with SqCC died shortly after fluid collection (range 2-313 days; mean: 58.1 days). CONCLUSIONS: Metastatic SqCC in serous effusions are rare, primarily arranged as single cells with rounded nuclei lacking visible nucleoli surrounded by a minimal amount of dense cytoplasm, and represent a dismal prognosis.

Predictive value of intra-abdominal lymph nodes in pancreatic endoscopic ultrasonography-guided fine-needle aspiration biopsy.

INTRODUCTION: Endoscopic ultrasonography (EUS)-guided fine-needle aspiration (FNA) biopsy is a commonly used method for the evaluation of pancreatic lesions. EUS-guided FNA of the intra-abdominal lymph nodes (LNs) can provide critical diagnostic information that is important for clinical management and tumor staging. This study examines the predictive value of intra-abdominal LN EUS-guided FNA biopsy associated with pancreatic lesions. MATERIALS AND METHODS: Over a 10-year period, the pathology database was searched for patients with concurrent pancreas and intra-abdominal LN EUS-guided FNA biopsy. The corresponding reports were reviewed, and clinical information and diagnostic results were recorded. RESULTS: There were 252 cases where both a pancreas lesion and intra-abdominal LN were biopsied. Of this group, 182 LNs were classified as negative (72%), 47 as positive (19%), and 23 as atypical (9%). Within the negative LN cohort, the pancreas FNAs fell into the following diagnostic categories: benign (47%), malignant (30%), and atypical/suspicious (23%). Within the positive LN cohort, the pancreas lesion correlated with the following diagnostic categories: malignant (89%), atypical (4%), and suspicious (6%). A positive LN EUS-guided FNA biopsy had a 98% positive predictive value for malignancy. Within the atypical LN cohort, the pancreas correlated with the following diagnostic categories: malignant (57%), atypical/suspicious (26%), and benign (17%). CONCLUSIONS: An atypical LN diagnostic category is strongly associated with a malignant pancreas lesion. A positive LN EUS-guided FNA biopsy has a 98% positive predictive value for pancreatic malignancy. A positive diagnostic category for an intra-abdominal LN can provide strong predictive evidence of a corresponding malignancy of the pancreas.

Florid diffuse peritoneal deciduosis mimicking carcinomatosis in a primigravida patient: a case report and review of the literature.

A case of a 27 year old G1P0 female with a dichorionic, diamniotic twin pregnancy presenting with premature rupture of membranes found to have omental caking and diffuse yellow-tan peritoneal nodules, clinically suspicious for carcinomatosis. The case work-up showed this to be an example of florid-diffuse peritoneal deciduosis mimicking carcinomatosis which has since resolved 4 months postpartum.

Clinicopathologic predictors of thyroid bed recurrence of differentiated thyroid cancer using ultrasound-guided fine-needle aspiration biopsies.

BACKGROUND: Monitoring changes in the thyroid bed (TB) is one of the clinical mainstays for surveillance of recurrent thyroid carcinoma. Fine-needle aspiration (FNA) is a diagnostic tool that is commonly used to aid in the identification of residual or recurrent disease. The aim of our study was to evaluate the efficacy of ultrasound-guided FNA of the TB in detecting recurrent thyroid cancer and to correlate the findings with clinicopathologic parameters to identify predictors of TB recurrence. METHODS: We retrieved cases of soft tissue masses within the TB that were evaluated for recurrence between January 1, 2006, and February 1, 2011. All ultrasound-guided FNA biopsies clinically suspected to indicate a lymph node metastasis and specimens with lymphocytes were excluded from the data. RESULTS: Of the 291 patients identified for evaluation of recurrence, 250 had papillary thyroid carcinoma (PTC), 10 had follicular carcinoma, 22 had medullary carcinoma, 7 had Hürthle cell carcinoma, and 2 had a previous thyroidectomy for an unknown type of thyroid carcinoma. For all FNAs that were clinically suspicious or intermediate for recurrence, the rate of positivity was 71.8% (209 patients). All cases diagnosed as "positive for PTC" or "suspicious for PTC" on TB FNA were found to have soft tissue metastasis on follow-up surgical resection. This resulted in a negative predictive value of 88.4% and a positive predictive value of 100%. The average time between thyroidectomy and TB FNA was 73.5 months. Of the patients with a previous diagnosis of PTC, those with suspicious/positive cytology were more likely to be women, to be older at thyroidectomy, to have documented metastasis to other sites as well as extrathyroidal extension and multifocal primary disease as compared with nondiagnostic/negative cytology cases. Patient age ≥45 years, primary tumor size at thyroidectomy, and surgical resection margin status had no statistical significance for predicting risk of TB recurrence. CONCLUSION: TB recurrence of PTC is most likely to occur in patients who have the following clinicopathologic parameters: documented metastasis to any site, extrathyroidal extension, and increased number of primary cancer foci.

Molecular markers for novel therapeutic strategies in pancreatic endocrine tumors.

OBJECTIVES: Pancreatic endocrine tumors (PETs) share numerous features with gastrointestinal neuroendocrine (carcinoid) tumors. Targets of novel therapeutic strategies previously assessed in carcinoid tumors were analyzed in PETs (44 cases). METHODS: Activating mutations in EGFR, KIT, and PDGFRA and nonresponse mutations in KRAS were evaluated. Copy number of EGFR and HER-2/neu was quantified by fluorescence in situ hybridization. Expression of EGFR, PDGFRA, VEGFR1, TGFBR1, Hsp90, SSTR2A, SSTR5, IGF1R, mTOR, and MGMT was measured immunohistochemically. RESULTS: Elevated EGFR copy number was found in 38% of cases but no KRAS nonresponse mutations. VEGFR1, TGFBR1, PDGFRA, SSTR5, SSTR2A, and IGF1R exhibited the highest levels of expression in the largest percentages of PETs.Anticancer drugs BMS-754807 (selective for IGF1R/IR), 17-(allylamino)-17-demethoxygeldanamycin (17-AAG, targeting Hsp90), and axitinib (directed toward VEGFR1-3/PDGFRA-B/KIT) induced growth inhibition of human QGP-1 PET cells with IC50 values (nM) of 273, 723, and 743, respectively. At growth-inhibiting concentrations, BMS-754807 inhibited IGF1R phosphorylation; 17-AAG induced loss of EGFR, IGF1R, and VEGFR2; and axitinib increased p21(CDKN1A) expression without inhibiting VEGFR2 phosphorylation. CONCLUSIONS: Results encourage further research into multidrug strategies incorporating inhibitors targeting IGF1R or Hsp90 and into studies of axitinib combined with conventional chemotherapeutics toxic to tumor cells in persistent growth arrest.

Molecular markers for novel therapies in neuroendocrine (carcinoid) tumors.

Neuroendocrine (carcinoid) tumors (NETs) are endocrine neoplasms occurring most frequently in gastrointestinal and bronchopulmonary (BP) systems. The majority of patients present with advanced disease for which few treatment options exist. We assessed 104 NETs (74 cases) for biomarkers targeted by anticancer drugs under development for other forms of cancer. Activating mutations were assessed in epidermal growth factor receptor (EGFR), stem cell factor receptor (KIT), and platelet-derived growth factor receptor alpha (PDGFRA), as well as non-response mutations in KRAS. Copy number of EGFR and HER-2/neu was quantified with fluorescence in situ hybridization. Immunohistochemical analyses were performed for EGFR, KIT, PDGFRA, somatostatin receptor subtypes 2A and 5 (SSTR5), vascular endothelial growth factor receptor 1, mammalian target of rapamycin (mTOR), insulin-like growth factor 1 receptor (IGF1R), heat shock protein 90 (Hsp90), and transforming growth factor-beta receptor 1 (TGFBR1). NETs lacked HER2-overexpression predictive of anti-HER2 response and KIT and PDGFRA activating mutations indicative of imatinib sensitivity. High EGFR aneusomy (20% of all cases) and elevated EGFR copy number (39%) were found, but few KRAS mutations associated with non-response to anti-EGFR therapy (3%). Hsp90, TGFBR1, IGF1R, and SSTR5 exhibited highest levels of immunohistochemical staining in the largest percents of tumors. In subsequent in vitro studies, anticancer drug 17-(allylamino)-17-demethoxygeldanamycin (17-AAG) (targeting Hsp90) inhibited proliferation of BP NET lines NCI-H727, NCI-H720, and NCI-H835 with IC(50) values of 70.4, 310, and 788 nM respectively; BMS-754807 (targeting IGF1R/IR) inhibited growth with IC(50) values of 428 nM, 2.8 microM, and 1 microM. At growth-inhibiting concentrations, 17-AAG (24 h) induced loss of EGFR and IGF1R in the IGF1R-expressing NCI-H727 line, and BMS-754807 (24 h) inhibited constitutive IGF1R autophosphorylation. Our results support further research into Hsp90, IGF1R, and EGFR as targets for developing new anticancer therapeutics for some NETs.