Augmented CPT1A Expression Is Associated with Proliferation and Colony Formation during Barrett's Tumorigenesis.

Obesity is a known risk factor for the development of gastroesophageal reflux disease (GERD), Barrett's Esophagus (BE) and the progression to esophageal adenocarcinoma. The mechanisms by which obesity contributes to GERD, BE and its progression are currently not well understood. Recently, changes in lipid metabolism especially in the context of a high fat diet have been linked to GERD and BE leading us to explore whether fatty acid oxidation plays a role in the disease progression from GERD to esophageal adenocarcinoma. To that end, we analyzed the expression of the rate-limiting enzyme, carnitine palmytoyltransferase 1A (CPT1A), in human tissues and cell lines representing different stages in the sequence from normal squamous esophagus to cancer. We determined uptake of palmitic acid, the most abundant fatty acid in human serum, with fluorescent dye-labeled lipids as well as functional consequences of stimulation with palmitic acid relevant to Barrett's tumorigenesis, e.g., proliferation, characteristics of stemness and IL8 mediated inflammatory signaling. We further employed different mouse models including a genetic model of Barrett's esophagus based on IL1ß overexpression in the presence and absence of a high fat diet and deoxycholic acid to physiologically mimic gastrointestinal reflux in the mice. Together, our data demonstrate that CPT1A is upregulated in Barrett's tumorigenesis and that experimental palmitic acid is delivered to mitochondria and associated with increased cell proliferation and stem cell marker expression.

Use of the Prostate Core Mitomic Test in Repeated Biopsy Decision-Making: Real-World Assessment of Clinical Utility in a Multicenter Patient Population.

BACKGROUND: Prostate cancer is the most common cancer diagnosed in men in developed countries. Using molecular testing may help to improve outcomes in this clinically challenging group. Since 2011, the Prostate Core Mitomic Test (PCMT), which quantifies a 3.4-kb mitochondrial DNA deletion strongly associated with prostate cancer, has been used by more than 50 urology practices accessing pathology services through our laboratory in New Jersey. However, the use of a molecular test can only be beneficial if it affects patient management and improves outcomes. OBJECTIVE: To determine whether repeated biopsy decision-making was affected in a quantifiable manner through the adjunct use of molecular testing with the PCMT. METHODS: In this observational study we conducted 2 independent, structured query language database queries of our patient records at our laboratory, QDx Pathology Services, in Cranford, NJ. Query 1 included all men who had a negative prostate biopsy and a negative PCMT between February 1, 2011, and June 30, 2013. Men with a previous diagnosis of cancer were excluded. Query 2 included all men who had a negative prostate biopsy and a repeated biopsy between February 1, 2011, and September 30, 2013. The data exported for each query included the unique specimen number for an index biopsy, the interval between biopsies where present, the unique specimen number for a follow-up biopsy where present, histopathology for all biopsies, the biopsy procedure dates, the patient's date of birth, and the PCMT result when utilized. The patient rebiopsy rates and intervals were compared between the patients who were using PCMT and those who were not to assess whether the adjunct use of the PCMT impacted the rebiopsy decision-making process. RESULTS: Query 1 identified 644 men who had a negative biopsy and a negative PCMT result within the study period. Query 2 identified 823 men with a repeat biopsy after the initial negative index biopsy within the study period. Of these men, 132 had PCMT to inform their care. This patient population of 1467 men originated from US-based clinical urology practices. Evaluation of the impact on physician behavior demonstrated a general trend toward the earlier detection of prostate cancer on repeat biopsy by an average of 2.5 months and a coincident increase in cancer detection rates for urologists using the deletion assay in their rebiopsy decision-making process. Importantly, this trend was only observed when men with atypical small acinar proliferation (ASAP) on index biopsy were not considered. In the 644 men with a negative PCMT result, only 35 (5.4%) were subjected to a follow-up biopsy, with 5 (14.3%) of the 35 men identified as having cancer. Finally, the cohort of 132 men who had PCMT and repeat biopsy was compared with the published data supporting PCMT's ability to predict rebiopsy outcome. The key metrics of sensitivity and negative predictive value were comparable and within the 95% confidence intervals of the reported work. CONCLUSION: Molecular tests, such as the PCMT, are useful in addressing the sampling error of prostate needle biopsy and providing additional evidence to inform the clinical uncertainty regarding initial negative prostate biopsy when ASAP is not present. Longitudinal monitoring of clinical impact indicators provides the necessary inputs to better allocation of healthcare resources in the short- and long-term.

HPV DNA testing in the triage of atypical squamous cells of undetermined significance (ASCUS): cost comparison of two methods.

Human papillomavirus (HPV) DNA testing for triage of cervical cytologies showing atypical squamous cells of undetermined significance (ASCUS) has become the standard of practice. Currently, Hybrid Capture II (HCII) is the preferred method for ASCUS triage. In situ hybridization for HPV represents an alternative to HCII and appears to have a superior specificity but is more expensive. We compare the reimbursement rates of ASCUS triage (HPV high risk) using the methods of HCII and INFORM (in situ hybridization for HPV) in a series of 431 ASCUS patients. The patients were followed for 1 yr, during which each patient had either colposcopic biopsy or follow-up cervical cytology after ASCUS HPV DNA triage. Eighty-nine patients were excluded from the analysis because of incomplete follow-up. The HPV triage percentages, colposcopic biopsy positivity rates and cervical cytology positivity percentages were calculated for each method. The reimbursement rates of the tests/procedures used in the analysis were those in effect at the University of Utah in 2003. The total triage and follow-up reimbursement costs were calculated for HCII and INFORM and compared.HCII referred 19.9% of patients to colposcopy, with a biopsy positivity rate of 25.6% for dysplasia. INFORM referred 11.8% of patients to colposcopy, of whom 34% had a biopsy diagnosis of dysplasia. HCII negative cases revealed 19% to have ASCUS or higher on the follow-up cervical cytology, while 19.9% of INFORM negative cases had a reading of ASCUS or higher at follow-up cytologic examination. The 1-yr HPV DNA triage and follow-up reimbursements for HCII were 316,942.00 US dollars per 1,000 women, and for the INFORM methodology, the reimbursements were 369,484.00 US dollars per 1,000 women. The INFORM method was associated with higher specificity and sent fewer (41%) patients to colposcopy than did HCII. Although this smaller referral rate reduced reimbursement costs associated with colposcopy, the increased reimbursement paid for follow-up cytologies and office visits of HPV DNA negative patient and the greater cost of the INFORM test results in higher overall reimbursement for INFORM. Based on these costs and diagnostic accuracies, it appears that the INFORM HPV technology represents a viable option to HCII ASCUS triage. INFORM HPV appears to be 16% more expensive than HCII but has the advantage of sending 41% fewer women to colposcopy.

HPV testing in liquid cytology specimens: comparison of analytic sensitivty and specificity for in situ hybridization and chemiluminescent nucleic acid testing.

OBJECTIVE: To compare the analytic sensitivity and specificity of Hybrid Capture 2 (HC2) (Digene Corporation, Gaithersburg, Maryland, U.S.A.) with in situ hybridization (ISH) in detecting high-risk types of HPV (HR-HPV). STUDY DESIGN: Performance characteristics of ISH and HC2 were compared in 99 consecutive cervical cytology samples diagnosed as low grade squamous intraepithelial lesion and processed by either the ThinPrep (Cytyc Corporation, Boxborough, Massachusetts, U.S.A.) or SurePath (TriPath Imaging Systems, Research Triangle Park, North Carolina, U.S.A.) method at 2 geographically different centers. Polymerase chain reaction (PCR) was used as the gold standard. RESULTS: Of the samples, 67% were positive for HR-HPV viral types by PCR. ISH had a sensitivity of 0.87 as compared to 0.95 (p = 0.11) for HC2. The specificity of ISH was significantly different from that of HC2 (0.57 vs. 0.13, respectively; p = 0.0004). The performance characteristics of ISH were not affected by the processing method or population tested. CONCLUSION: The sensitivity of ISH is comparable to that of HC2, with significantly superior specificity, and is therefore an efficacious alternative to HC2 for triaging patients with abnormal cervical cytology results.

Role of HPV DNA testing in predicting cervical intraepithelial lesions: comparison of HC HPV and ISH HPV.

Human papillomavirus (HPV) is widely accepted as the primary agent involved in the development of squamous intraepithelial neoplasia and cervical carcinoma. Several commercial tests are available for detecting HPV DNA. This study compares the efficacy of INFORM HPV (in situ hybridization [ISH] HPV) and HCII (HC HPV) in predicting cervical lesions. A total of 762 sequential Papanicolaou (Pap) smears determined by cytologic examination to be either atypical squamous cells of undetermined significance (ASC-US) or low-grade squamous intraepithelial lesions (LSIL) were tested by both Hybrid Capture (HC) HPV and ISH HPV; 250 follow-up biopsies were reviewed as the reference standard for presence or absence of a lesion. ISH HPV and HC HPV differed significantly in accurately predicting biopsy findings from ASC-US and LSIL cases. The overall sensitivity and specificity of ISH HPV were 97% (28/29) and 86% (191/221); and HC HPV was 79% (23/29) and 56% (123/221). The positive predictive value (PPV) of ISH HPV was 48% (28/58) vs HC HPV value of 19% (23/121). Negative predictive value (NPV) was also better with ISH HPV at 99% (191/192) and HC HPV at 95% (123/129). Of equal importance, ISH HPV demonstrated a lower false-positive rate compared to HC HPV, 12% (30/250) vs 39% (98/250), as well as having a slightly lower false-negative rate 0.4% (1/250) vs 2.4% (6/250). ISH HPV is more predictive of biopsy histopathology in patients with detectable cervical lesions than is HC HPV. Effective triage of patients by HPV analysis using ISH HPV as compared to HC HPV has the potential of significant public health impact by reducing unnecessary colposcopies, as well as adverse medical, social, and psychological patient consequences.