Chengquan Zhao

Identification of the most sensitive and robust immunohistochemical markers in different categories of ovarian sex cord-stromal tumors.

Different immunohistochemical sex cord-stromal markers have been previously studied in various types of ovarian sex cord-stromal tumors; however, the sensitivity for sex cord-stromal lineage may vary between markers, and some markers may not be as sensitive in some types of sex cord-stromal tumors compared with other tumors in this spectrum of neoplasms. The goals of this study were to determine which immunohistochemical markers are the most sensitive and immunohistochemically robust for sex cord-stromal lineage within a given type of ovarian sex cord-stromal tumor, and to establish whether there are substantial differences of expression of these markers between different types of sex cord-stromal tumors. Immunohistochemical stains for markers which have known variable specificity for sex cord-stromal lineage [inhibin, calretinin, MART-1/melan-A, CD99, steroidogenic factor 1 (SF-1, adrenal 4-binding protein), and WT1], were performed in 127 cases of 5 different types of ovarian sex cord-stromal tumors: adult granulosa cell tumor (n=32), Sertoli cell tumor (n=27), Sertoli-Leydig cell tumor (n=18), steroid cell tumor (n=25), and fibroma/fibrothecoma (n=25). All cases in each type of sex cord-stromal tumor expressed SF-1. Inhibin and calretinin were expressed in all groups of tumors but with a lesser frequency (56% to 100% and 36% to 100% of cases, respectively). All types of tumors except steroid cell tumor expressed WT1. Fibroma/fibrothecoma was the only type of tumor that did not express CD99. The only tumor groups that showed expression of MART-1 were Sertoli-Leydig cell tumor (restricted to the Leydig cell component) and steroid cell tumor (94% and 96% of cases, respectively). The type of sex cord-stromal tumor that was least frequently positive for several of the different markers studied was fibroma/fibrothecoma. Among all tumor groups combined, inhibin and WT1 were the 2 markers showing the most diffuse expression. Likewise, the single marker showing the most optimal combination of diffuse and strong staining (immunohistochemical composite score: possible range, 1 to 12) varied between tumors: adult granulosa cell tumor-inhibin (score 10.0); Sertoli cell tumor-WT1 (score 10.8); Sertoli-Leydig cell tumor (Sertoli cell component)-WT1 (score 10.4); steroid cell tumor-inhibin (score 11.2); and fibroma/fibrothecoma-WT1 (score 8.9). We conclude that most immunohistochemical sex cord-stromal markers have sufficient sensitivity for sex cord-stromal lineage. Although each of the different types of sex cord-stromal tumors has a slightly unique immunoprofile in terms of frequency and extent of expression, these differences are relatively minor for most types of tumors with certain exceptions (eg, WT1 is not diagnostically useful in steroid cell tumor; CD99 is not diagnostically useful in fibroma/fibrothecoma; the only sex cord-stromal tumor for which MART-1 is diagnostically useful is steroid cell tumor; inhibin and calretinin are less diagnostically useful in fibroma/fibrothecoma than in the other types of tumors, but expression in fibrothecoma was higher than in fibroma). SF-1 is the most sensitive sex cord-stromal marker among the most common types of sex cord-stromal tumors. Given the findings relating to sensitivity and extent of expression in this study, and known specificity in the literature, the most informative sex cord-stromal markers to be used for the distinction from nonsex cord-stromal tumors are inhibin, calretinin, SF-1, and WT1 (the exact number of markers to be used should be based on the degree of difficulty of the case and level of experience of the pathologist); however, the utility of immunohistochemistry for the diagnosis of fibroma/fibrothecoma is somewhat limited.

Histologic follow-up results in 662 patients with Pap test findings of atypical glandular cells: Results from a large academic womens hospital laboratory employing sensitive screening methods

OBJECTIVE Atypical glandular cell (AGC) Pap interpretations and screening for glandular neoplasias remain major challenges. We document the largest reported AGC histopathologic follow-up experience and include verification bias-adjusted data on laboratory screening sensitivity. METHODS AGC Pap tests of endocervical origin (AGC-EC), endometrial origin (AGC-EM), and not otherwise specified (AGC-NOS) were documented at a center serving an older low risk population. 98% of Pap tests were liquid-based cytology (LBC) specimens screened using computer-assisted screening. Follow-up diagnoses were correlated with cytology and stratified into age groups. Screening sensitivity was assessed by examining Pap results during 1 year preceding neoplastic diagnoses. Verification bias was adjusted with findings in over 2000 patients with hysterectomies. RESULTS Of 247,131 Pap tests, 1021 (0.41%) reported AGC results and 662 cases had tissue follow-up. Precancerous or malignant neoplastic histologic outcomes were documented in 101 patients (15.3%), including 8.3% cervical, 6.3% endometrial, and 0.6% ovarian. AGC results were most often associated with neoplastic cervical outcomes in women younger than 40 and with neoplastic endometrial outcomes in women 50 or older. AGC-NOS with a squamous cell abnormality and AGC-EC results suggested cervical neoplasia, while AGC-EM results suggested endometrial neoplasia. CONCLUSIONS AGC Pap results detected significant numbers of cervical and non-cervical neoplasias. Since 38 of 44 (86%) of AGC-detected carcinomas were endometrial or ovarian, HPV co-testing would not have aided screening in detecting the majority of malignancies diagnosed after AGC Pap results. Verification bias-adjusted Pap screening sensitivity in the laboratory for detection of significant neoplastic cervical disease was 93%.

Adjunctive Human Papillomavirus DNA Testing Is a Useful Option in Some Clinical Settings for Disease Risk Assessment and Triage of Females With ASC-H Papanicolaou Test Results

CONTEXT Recent guidelines recommend colposcopy for women with atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H). objective: To determine whether adjunctive high-risk human papillomavirus (hrHPV) testing is useful for disease risk assessment in females with ASC-H Papanicolaou (Pap) test results. DESIGN High-risk HPV prevalence and histopathologic follow-up data on 1187 females with ASC-H ThinPrep Pap test (TPPT) and hrHPV test results were analyzed. RESULTS ASC-H was reported in 1646 (0.59%) [corrected] of 277 400 (270 338 TPPT and 7062 conventional) Pap test results. The difference in ASC-H detection rates between TPPTs and conventional Pap smears was statistically significant (0.60% vs 0.38%; P = .02). High-risk HPV was detected in 589 (49.6%) of 1187 females with ASC-H TPPT and hrHPV testing. The hrHPV DNA-positive rate in females younger than 40 years was 54.7%, significantly higher than the 36.5% in women 40 years and older. Among 505 females with histopathologic follow-up, cervical intraepithelial neoplasia 2/3 was identified in 32.7% of hrHPV-positive females compared with 1.2% in hrHPV-negative females. The sensitivity, specificity, positive predictive value, and negative predictive value of ASC-H cytology in conjunction with hrHPV DNA testing results for detection of cervical intraepithelial neoplasia 2/3 were 96.1% versus 100.0%, 54.0% versus 68.4%, 35.8% versus 20.8%, and 98.1% versus 100.0% in females younger than 40 years and women 40 years and older, respectively. CONCLUSIONS Our data suggest that reflex hrHPV testing is a highly useful option for women with ASC-H Pap tests. Females with ASC-H and negative hrHPV testing may be more efficiently managed by follow-up with regular Pap and hrHPV testing rather than universal colposcopy, especially for women 40 years and older.

Sf-1 is a Diagnostically Useful Immunohistochemical Marker and Comparable to Other Sex Cord-stromal Tumor Markers for the Differential Diagnosis of Ovarian Sertoli Cell Tumor

Immunohistochemistry can be an important part of the diagnosis of Sertoli cell tumor of the ovary, including distinction from non-sex cord-stromal tumors such as the sertoliform variant of endometrioid carcinoma and carcinoid. Several good markers for this differential diagnosis have been identified, particularly inhibin, Wilms tumor 1 gene product (WT1), epithelial membrane antigen, and chromogranin; however, many available markers have limitations to some degree. Steroidogenic factor 1 (SF-1; adrenal 4-binding protein; Ad4BP) is a nuclear transcription factor involved in gonadal and adrenal development. In the testes, SF-1 is expressed in Sertoli cells. Immunohistochemical expression of this marker in ovarian sex cord-stromal tumors, including utility for differential diagnosis, has not been rigorously evaluated. As an extension of our previous immunohistochemical studies of ovarian Sertoli cell tumor, expression of SF-1 and comparison with WT1 and inhibin were assessed in 111 primary ovarian tumors: 27 Sertoli cell tumors, 60 endometrioid tumors (including borderline tumors, conventional well-differentiated carcinomas, and sertoliform variants of carcinoma), and 24 carcinoids. SF-1 was expressed in 100% of Sertoli cell tumors but not in endometrioid tumors or carcinoid. WT1 was expressed in 100% of Sertoli cell tumors and 17% of endometrioid tumors; all carcinoids were negative. Inhibin was expressed in 96% of Sertoli cell tumors and 2% of endometrioid tumors (4% of conventional well-differentiated carcinomas); all carcinoids were negative. The extent of expression of all 3 markers was similar in Sertoli cell tumor but greatest for WT1: 63%, 96%, and 78% of cases showed expression of SF-1, WT1, and inhibin, respectively, in more than 50% of tumor cells. Immunohistochemical composite scores combining both extent and intensity of staining in positive cases were calculated for Sertoli cell tumor (possible range: 1–12). Combined extent/intensity of immunostaining was similar for all 3 markers, but WT1 showed the most robust immunoreactivity in positive cases (mean immunohistochemical composite scores for SF-1, WT1, and inhibin: 6.1, 10.8, and 7.8, respectively). We conclude that for the differential diagnosis with endometrioid tumors and carcinoid of the ovary, SF-1 is a sensitive and specific immunohistochemical marker for Sertoli cell tumor and that SF-1 is diagnostically comparable with other good sex cord-stromal markers.

Very low human papillomavirus DNA prevalence in mature women with negative computer-imaged liquid-based Pap tests

The prevalence of high‐risk Human Papillomavirus DNA (hrHPV DNA) in women with negative Papanicolaou (Pap) test results provides a measure of residual risk for cervical neoplasia after cytology screening. The purpose of this study was to document the prevalence of hrHPV DNA in several thousand women ages ≥30 years with negative ThinPrep Imaging System (TIS)‐imaged Pap test results in a large academic hospital cytology laboratory.

Clinical utility of adjunctive high-risk human papillomavirus DNA testing in women with Papanicolaou test findings of atypical glandular cells.

CONTEXT Atypical glandular cell (AGC) Papanicolaou (Pap) test interpretations are challenging. Most biopsy findings are benign, but AGC results may also reflect highly significant noninvasive neoplastic and malignant histologic outcomes. High-risk human papillomavirus (hrHPV) test use with AGC Pap test results is evolving. OBJECTIVE To further evaluate the utility and limitations of hrHPV testing with AGC Pap tests. DESIGN Hospital records were searched for AGC Pap tests results from June 1, 2005, to August 31, 2007. Cases of AGC with hrHPV tests and histopathologic follow-up were included. RESULTS Of the 662 women with AGC Pap test results and follow-up analyzed, hrHPV results were available for 309 (46.7%) and were positive in 75 cases (24.3%). Among the 75 cases with hrHPV+ AGC results, 13 (17.3%) had cervical intraepithelial neoplasia grades 2/3, 10 (13.3%) had adenocarcinoma in situ, and 3 (4.0%) had cervical invasive adenocarcinoma, whereas for 234 women with hrHPV(-) results, 1 (0.4%) had cervical intraepithelial neoplasia grades 2/3, 1 (0.4%) had adenocarcinoma in situ, 1 each (0.4%) had cervical adenocarcinoma and ovarian carcinoma, and 8 (3.4%) had endometrial carcinoma. CONCLUSIONS Positive hrHPV AGC results were most strongly associated with cervical intraepithelial neoplasia grades 2/3 and adenocarcinoma in situ in women younger than 50 years. Positive hrHPV AGC results were also present in all 3 cases of invasive cervical adenocarcinoma in women younger than 50 years. Of note, hrHPV(-) AGC results were present in 10 of 13 carcinomas (76.9%) detected after AGC Pap tests, all in women 40 years or older with endometrial adenocarcinomas (n = 8), ovarian carcinoma (n = 1), and cervical adenosquamous carcinoma in a woman (n = 1) in her 50s. Testing for hrHPV after AGC Pap testing was most helpful in the detection of cervical intraepithelial neoplasia grades 2/3, adenocarcinoma in situ, and invasive cervical adenocarcinomas in women younger than 50 years.

High-risk HPV DNA detected in less than 2% of over 25,000 cytology negative imaged liquid-based Pap test samples from women 30 and older

OBJECTIVE The purpose of this study was to document the prevalence of high-risk HPV DNA (HPV) in the largest cohort of woman studied to date with negative ThinPrep Imaging system (TIS)-imaged Pap tests. METHODS Women with negative (TIS)-imaged ThinPrep Pap Tests (TPPT) who also were tested for HPV were identified between July 1, 2005 and December 31, 2007 from a large womens hospital practice. HPV detection rates were compared for women with either presence or absence of a transformation zone/endocervical cell sample (EC/TZS). RESULTS 26,558 negative TPPT also underwent HPV testing. HPV detection was higher in women younger than 30 and sharply declined in women 30-39 (P<0.001). Declining HPV detection rates continued in the 40-49 age group (age 30-39 vs. 40-49; 2.8% vs. 1.7%, P<0.001) and then levelled off. No statistically significant difference for HPV prevalence was identified comparing women with and without a TZ/ECS. CONCLUSION This is the largest study to date documenting very low HPV detection rates in women screened cytology negative with computer-imaged liquid-based Pap methods now representing a major portion of the U.S. cervical cytology market. Findings of very low rates of HPV detection in 490 (1.9%) of 25,259 cytology negative women 30 and older extend and confirm previously reported findings in smaller study populations. Because HPV testing provides an objective measure of relative residual risk for cervical neoplasia after screening, these data are relevant to discussions on how best to combine cytology and HPV testing in screening low risk populations.

Clinical Significance of Atypical Glandular Cells in Conventional Pap Smears in a Large, High-Risk U.S. West Coast Minority Population

OBJECTIVE To determine the incidence of clinically significant lesions on subsequent histologic follow-up in high-risk, predominantly minority patients with atypical glandular cells (AGC). STUDY DESIGN A retrospective study was done on conventional Pap smears diagnosed as AGC of endocervical origin (AGC-EC), AGC of endometrial origin (AGC-EM) and AGC not otherwise specified (AGC-NOS) between January 1, 2003, and December 31, 2005. Histologic diagnoses were correlated with cytologic diagnoses. RESULT Confirmed AGC cases were divided into 4 categories: 187 AGC-NOS, 169 AGC-EC, 68 AGC and atypical squamous cells of undetermined significance (ASCUS) and 36 AGC-EM. A total of 105 patients (22.8%) had significant precancerous (cervical intraepithelial neoplasia [CIN] 2/3, adenocarcinoma in situ [AIS]) or malignant (carcinoma) histologic outcomes. CIN 2/3 was the most common significant histologic outcome in women with AGC and ASCUS and patients <35 years with AGC. Endometrial neoplasia was the most common significant outcome in women with AGC-NOS and AGC-EM Pap results and in AGC patients > or =35. In women with AGC-EC Pap results, glandular cervical neoplasia occurred in 8.3% and CIN 2/3 in 5.9% offollow-up biopsies. CONCLUSION AGC subtype and age significantly affect the probability of precancerous and malignant follow-up findings and anatomic site of neoplastic lesions. Access to newer screening technologies such as high-risk HPVDNA testing and liquid-based cytology will likely benefit such high-risk populations.

The cytologic criteria of malignancy

Cytology and cell biology are two separate fields that share a focus on cancer. Cancer is still diagnosed based on morphology, and surprisingly little is known about the molecular basis of the defining structural features. Cytology uses the smallest possible biopsy for diagnosis by reducing morphologic “criteria of malignancy” to the smallest scale. To begin to develop common ground, members of the American Society of Cytopathology Cell Biology Liaison Working Group classify some of the “criteria of malignancy” and review their relation to current cell biology concepts. The criteria of malignancy are extremely varied, apparently reflecting many different pathophysiologies in specific microenvironments. Criteria in Group 1 comprise tissue‐level alterations that appear to relate to resistance to anoikis, alterations in cell adhesion molecules, and loss of apical–basal polarity. Criteria in Group 2 reflect genetic instability, including chromosomal and possibly epigenetic instability. Criteria in Groups 3 are subcellular structural changes involving cytoplasmic components, nuclear lamina, chromatin and nucleoli that cannot be accounted for by genetic instability. Some distinct criteria in Group 3 are known to be induced by cancer genes, but their precise structural basis remains obscure. The criteria of malignancy are not closely related to the histogenetic classification of cancers, and they appear to provide an alternative, biologically relevant framework for establishing common ground between cytologists and cell biologists. To understand the criteria of malignancy at a molecular level would improve diagnosis, and likely point to novel cell physiologies that are not encompassed by current cell biology concepts. J. Cell. Biochem. 110: 795–811, 2010.

Screening Test Results Associated With Cancer Diagnoses in 287 Women With Cervical Squamous Cell Carcinoma

CONTEXT The limitations of newer cervical screening tests are still being studied. OBJECTIVE To investigate those limitations, we conducted a retrospective review of 287 cases of cervical squamous cell carcinoma (SCC). DESIGN A search through 5 years of records identified 287 women with cervical SCC diagnoses. Clinical history, histopathology, and liquid-based cytology (ThinPrep) and Hybrid Capture 2 (HC2) high-risk human papillomavirus (HPV) results were documented. Polymerase chain reaction HPV tests were performed on SCC tissues with prior negative HC2 results. RESULTS Of 287 women, 156 (54.4%) had abnormal cytology results, and 75 (26.1%) had abnormal clinical findings triggering tissue diagnoses of SCC. Among 156 patients with abnormal Papanicolaou (Pap) test results, more-seriously abnormal Pap test results were reported in 142 women (91.1%). Among 31 women with Pap and HC2 HPV cotesting within 1 year of SCC diagnoses, 28 (90%) were Pap⁺/HPV⁺, 2 (15.5%) were Pap⁺/HPV⁻, and 1 (3%) was Pap⁻/HPV⁻. Two of 3 women with negative HC2 results before SCC diagnosis had abnormal Pap results; 1 had negative Pap reports with questionable lesional cells mimicking atrophy. In all 3 cases of SCC with negative HC2 results, HPV-18 was detected; in 2 cases (66%), HPV-16 was also detected. CONCLUSIONS Although abnormal cytology preceded most SCC diagnoses, about one-third of patients were referred for diagnostic testing because of clinical suspicion alone. Among 31 SCC cases with liquid-based cytology and high-risk HPV cotest results less than 1 year before SCC diagnoses, 2 patients (6.5%) had Pap⁺/HPV⁻ results, and 1 patient (3.2%) had Pap⁻/HPV⁻ results. Polymerase chain reaction detected high-risk HPV DNA in tumor tissues of 3 SCC cases with recent HC2⁻ results.