CDK9 Expression Shows Role as a Potential Prognostic Biomarker in Breast Cancer Patients Who Fail to Achieve Pathologic Complete Response after Neoadjuvant Chemotherapy.

Failure to achieve pathologic complete response is associated with poor prognosis in breast cancer patients following neoadjuvant chemotherapy (NACT). However, prognostic biomarkers for clinical outcome are unclear in this patient population. Cyclin-dependent kinase 9 (CDK9) is often dysregulated in breast cancer, and its deficiency results in genomic instability. We reviewed the records of 84 breast cancer patients from Emory University's Winship Cancer Institute who had undergone surgical resection after NACT and had tissue available for tissue microarray analysis (TMA). Data recorded included disease presentation, treatment, pathologic response, overall survival (OS), locoregional recurrence free survival (LRRFS), distant-failure free survival (DFFS), recurrence-free survival (RFS), and event-free survival (EFS). Immunohistochemistry was performed on patient samples to determine CDK9 expression levels after NACT. Protein expression was linked with clinical data to determine significance. In a Cox proportional hazards model, using a time-dependent covariate to evaluate the risk of death between groups beyond 3 years, high CDK9 expression was significantly associated with an increase in OS (HR: 0.26, 95% CI: 0.07-0.98, p=0.046). However, Kaplan-Meier curves for OS, LRRFS, DFFS, RFS, and EFS did not reach statistical significance. The results of this study indicate that CDK9 may have a potential role as a prognostic biomarker in patients with breast cancer following NACT. However, further validation studies with increased sample sizes are needed to help elucidate the prognostic role for CDK9 in the management of these patients.

Relationship of histologic grade and histologic subtype with oncotype Dx recurrence score; retrospective review of 863 breast cancer oncotype Dx results.

PURPOSE: Oncotype Dx (ODx) is a multigene assay that is prognostic and predictive in estrogen receptor (ER) positive early breast cancer. ODx recurrence score (RS) is reported to be histologic grade dependent. Relationship of RS with breast cancer histologic subtypes is unknown. This study was designed to assess the relationship of histologic subtype with RS. Histologic grade dependence of RS was also investigated. METHODS: Results of consecutive ODx tests (1/2007-7/2016) from two institutions were reviewed. Histologic subtypes (in: Lakhani et al., WHO classification, IARC Press, Lyon, 2012), combined Nottingham histologic grade, age and tumor size were recorded from pathology reports. Univariate and multivariate analysis was performed to investigate the relationship between RS and ODx risk categories and histologic subtypes, grade, age and tumor size. RESULTS: RS was grade dependent. RS of grade 1 and grade 2 tumors were significantly lower than grade 3 tumors. There was no high-risk grade 1 tumor. In favorable histologic subtypes there was no high-risk tumor. Mean RS of grade 1 lobular tumors was significantly higher than grade 1 ductal tumors. Using newer ODx cut-offs, 5 grade 1 tumors were reclassified as high risk (RS > 25) and grade 3 lobular tumors showed significantly higher rate of reclassification as high-risk than grade 3 ductal tumors. In a multivariate analysis, only grade showed a significant positive correlation with RS. Adding dichotomous histologic subtyping (favorable vs. non-favorable) to grade further improved correlation with RS. CONCLUSIONS: The Oncotype Dx result is impacted by histologic grade and histologic subtype. Tumors with favorable histologic subtypes and histologic grade 1 tumors do not have high-risk RS. High RS in a grade 1 tumor or in a tumor with favorable histology is unusual that warrants further investigation. Invasive lobular carcinomas rarely show high-risk RS. Histologic grade and histologic subtype should be considered while ordering ODx testing.

Cytology as a screening tool for anal squamous intraepithelial lesion for HIV positive men: 10-year experience in an inner city hospital.

INTRODUCTION: Human papillomavirus (HPV) and anal carcinoma are prevalent in high-risk patients including human immunodeficiency virus (HIV)-positive patients. There are currently no clear guidelines for screening, however. We assessed anal cytology specimens and HPV testing at an inner-city hospital by correlating anal cytology with anal biopsy (bx), and evaluated if results differed with traditional proctoscopy (TP) or high-resolution anoscopy (HRA). MATERIALS AND METHODS: 209 anal cytology and subsequent biopsies taken during the period 2003-2014 from 152 male patients were reviewed. Demographic data for age, sex, HIV, HPV, cytology, histology, and the method of biopsy were analyzed. RESULTS: All specimens were followed by a biopsy within a period of 6 months. Ninety-seven percent of patients were HIV-positive and 43% had AIDS. Lesions most diagnosed on cytology were low-grade squamous intraepithelial lesion (LSIL) (52%) and atypical squamous cells of undetermined significance (ASC-US) (21.5%). Lesions most diagnosed on bx were anal intraepithelial neoplasia (AIN) grade 2-3 (52%) and AIN grade 1 (37%). Almost all ASC-US cases tested for HPV were positive (97%). There was cytology histology correlation in 48% of LSIL and 83% of high-grade squamous intraepithelial lesions. Anal cytology had 97% sensitivity in detecting AIN and carcinoma and a positive predictive value of 96%. There was no difference in rate of detection of AIN 1and AIN 2-3 on bx using TP versus HRA. CONCLUSION: Screening in high-risk patients detected almost all high- and low-grade squamous intraepithelial lesions, however, anal cytology alone could not predict the degree of dysplasia. It may be prudent to perform anal bx in all atypical anal cytology. Clear guidelines are needed for screening of a high risk population.

A gemcitabine sensitivity screen identifies a role for NEK9 in the replication stress response.

The Replication Stress Response (RSR) is a signaling network that recognizes challenges to DNA replication and coordinates diverse DNA repair and cell-cycle checkpoint pathways. Gemcitabine is a nucleoside analogue that causes cytotoxicity by inducing DNA replication blocks. Using a synthetic lethal screen of a RNAi library of nuclear enzymes to identify genes that when silenced cause gemcitabine sensitization or resistance in human triple-negative breast cancer cells, we identified NIMA (never in mitosis gene A)-related kinase 9 (NEK9) as a key component of the RSR. NEK9 depletion in cells leads to replication stress hypersensitivity, spontaneous accumulation of DNA damage and RPA70 foci, and an impairment in recovery from replication arrest. NEK9 protein levels also increase in response to replication stress. NEK9 complexes with CHK1, and moreover, NEK9 depletion impairs CHK1 autophosphorylation and kinase activity in response to replication stress. Thus, NEK9 is a critical component of the RSR that promotes CHK1 activity, maintaining genome integrity following challenges to DNA replication.

Incidental gynecologic neoplasms in morcellated uterine specimens: a case series with follow-up.

Laparoscopic hysterectomy with morcellation (LHM) is considered a safe and less invasive alternative to other hysterectomy techniques by shortening postoperative hospital stay and patient recovery. Sparse incidental gynecologic neoplasms after LHM have been reported; however, the frequency and subsequent follow-up have not been systematically investigated in a large case series. We aimed to determine the frequency and types of incidental findings after LHM with clinical outcomes. An electronic chart review was conducted searching all cases of LHM performed within 5 years to determine the incidence of unexpected gynecologic neoplasms and subsequent peritoneal disease. Patient demographics, prior preoperative investigation, and subsequent follow-up were investigated. For comparison, the overall frequency of pertinent uterine neoplasms was noted during the study period. Of the 352 cases of LHM identified, 3 harbored unsuspected malignancies, an incidence of 0.9%. Four variant smooth muscle tumors (1.1%) and 5 benign non-smooth muscle neoplasms (1.4%) were identified at the time of initial morcellation. Two cases of subsequent peritoneal "implanted" leiomyoma were identified (0.6%). Of malignant or atypical mesenchymal neoplasms diagnosed at our institution during the study period, 8.6% were diagnosed in a morcellated specimen. There is a clinically important risk of occult malignant or atypical neoplasms in morcellated uterine specimens. Proper pathologic evaluation of malignant or atypical uterine neoplasms is limited when a uterus is morcellated. Patients undergoing morcellation procedures are also potentially at risk for dissemination of disease. Clinicians and patients should be aware of these risks when discussing surgical options for hysterectomy.

Are biosafety practices in anatomical laboratories sufficient? A survey of practices and review of current guidelines.

Biosafety practices in anatomical pathology laboratories are crucial to prevent unnecessary exposures to both chemical and biological agents. Regulatory and guidance agencies have general regulations and recommendations regarding anatomical pathology laboratory biosafety practices. This study aimed to determine if professionals' perceptions and actual practice mirror these guidelines. Current available regulations and recommendations for biosafety practices in anatomical pathology laboratories were reviewed and used to construct a brief, validated online survey distributed to anatomical pathology professionals. The survey was completed by 39 survey participants in pathology departments from diverse institutions. An average of 44% of respondents reported receiving inadequate biosafety training. At survey initiation, 61.5% of respondents felt that the risks of chemical and infectious disease exposures had been clearly explained to them; however, by completion of the survey, only 21% believed risks to be clearly explained. Respondents use a variety of personal protective equipment, yet only 60% would have been classified as meeting recommendations. Most respondents reported having a needle stick or cut (56.3%) or formalin exposure by splash or prolonged direct skin contact (62.5%). The survey indicated that there is a dire need for improved training in anatomical pathology biosafety as daily practices do not reflect current guidelines. In addition, improved training on exposure risks, including needle-stick injuries, personal protective equipment, and chemical hazards, is needed. Finally, the success of this training should be monitored locally as regulatory agency requirements do not seem to alter daily practice.

Triple negative breast cancer is associated with an increased risk of residual invasive carcinoma after lumpectomy.

BACKGROUND: To assess the potential mechanisms that may underlie increased local failure in triple negative (TN) breast cancers, an analysis was performed of the risk of residual carcinoma after lumpectomy with correlation to pathologic factors, including molecular phenotype. METHODS: A review of pathologic specimens was performed for women with invasive breast cancer treated with lumpectomy followed by reexcision. Data were collected on age; tumor size, grade, and nodal stage; estrogen receptor, progesterone receptor, and human endothelial growth factor receptor 2 (Her2); extensive intraductal component; lymphovascular invasion; margins; and reexcision findings. Univariate and multivariate logistic regression analyses were performed to evaluate for associations between pathologic features of the lumpectomy specimen and reexcision findings. Molecular phenotypes were defined by conventionally used immunohistochemical pattern. RESULTS: Data were collected on 369 patients with breast cancer. The median age was 57 years, median tumor size was 1.5 cm, 36% had positive margins, 32% had positive lymph nodes, 73.5% had the luminal A subtype, 9.5% had the luminal B subtype, 4.5% were Her2-enriched, and 12.5% were TN. Overall, 32% of patients had invasive cancer in their reexcision specimens, and 51% of those with the TN subtype had residual invasive disease on reexcision compared with 30% to 31% for other subtypes. On univariate analysis, age, tumor size, margin status, lymphovascular invasion, nodal status, and TN subtype were associated with elevated risk of residual invasive cancer. On multivariate analysis using a forward stepwise model, TN subtype maintained significance, with an odds ratio of 3.28 (P = .002). CONCLUSION: TN subtype has a statistically significant association with an increased risk of residual tumor. This suggests the putative increase in the risk of local failure in TN patients may be related to increased residual tumor burden.