Debbi H. Conlon

Tumor infiltrating Foxp3+ regulatory T-cells are associated with recurrence in pathologic stage I NSCLC patients

Early stage lung cancer has a variable prognosis, and there are currently no markers that predict which patients will recur. This study examined the relation between tumor‐regulatory T (Treg) cells and total tumor‐infiltrating T‐cell lymphocytes (TIL) to determine whether they correlated with recurrence.

High Gene Expression of TS1, GSTP1, and ERCC1 Are Risk Factors for Survival in Patients Treated with Trimodality Therapy for Esophageal Cancer

Purpose: To assess the relationship between molecular markers associated with chemotherapy resistance and survival in esophageal cancer patients treated with trimodality therapy. Experimental Design: The original pretreatment formalin-fixed, paraffin-embedded endoscopic esophageal tumor biopsy material was obtained from 99 patients treated with concurrent cisplatin plus 5-fluorouracil plus 45 Gy radiation followed by resection at Duke University Medical Center (Durham, NC) from 1986 to 1997. cDNA was derived from the biopsy and analyzed to determine mRNA expression relative to an internal reference gene (β-actin) using fluorescence-based, real-time reverse transcription-PCR. Possible markers of platinum chemotherapy association [glutathione S-transferase π (GSTP1) and excision cross-complementing gene 1 (ERCC1)] and 5-fluorouracil association [thymidylate synthase 1 (TS1)] were measured. Results: Cox proportional hazards model revealed a significant inverse, linear effect for TS1 with respect to survival (P = 0.007). An inverse relationship between TS1 expression and treatment response was also detected (P ≤ 0.001). Univariate analysis identified an association with decreased survival for GSTP1 ≥ 3.0 (P = 0.05). In multivariate analyses, TS1 >6.0, ERCC1 >3, and GSTP1 >3 were statistically significant predictors of decreased survival (P = 0.007). Additionally, the presence of ERCC1 >3.0 or TS1 >6.0 was associated with an ∼2-fold increase in the risk of cancer recurrence (P = 0.086 and 0.003, respectively). Conclusion: The measurement of relative gene expression of molecular markers associated with chemoresistance in endoscopic esophageal tumor biopsies may be a useful tool in assessing outcome in patients with trimodality-treated esophageal cancer. These data should be validated further in larger prospective studies.

Predicting the Sites of Metastases From Lung Cancer Using Molecular Biologic Markers

BACKGROUND The use of molecular markers in staging non-small cell lung cancer (NSCLC) has been supported in retrospective prognostic models but has not been evaluated in predicting sites of metastases. METHODS Pathologic specimens were collected from 202 patients after complete resection for stage I NSCLC, who were subsequently found to have no metastases at 5 years (n = 108), isolated brain metastases (n = 25), or other distant metastases (n = 69). A panel of eight molecular markers of metastatic potential was chosen for immunohistochemical analysis of the tumor: p53, erbB2, angiogenesis factor viii, EphA2, E-cadherin, urokinase plasminogen activator (UPA), UPA receptor, and plasminogen activator inhibitor. RESULTS Patients with isolated brain relapse had significantly higher expression of p53 (p = 0.02) and UPA (p = 0.002). The quantitative expression of E-cadherin was used to predict the site of metastases using recursive partitioning: 0 of 92 patients with E-cadherin expression of 0, 1, or 2 developed isolated cerebral metastases; 0 of 33 patients with E-cadherin expression of 3 with UPA of 1 or 2 and ErbB2 of 0 developed brain metastases. Of the remaining patients at risk (UPA = 3), the risk of isolated cerebral metastases was 21 of 57 patients (37%). CONCLUSIONS This study demonstrates that molecular markers may predict the site of relapse in early stage NSCLC. If validated in an ongoing prospective study, these results could be used to select patients with isolated brain metastases for adjuvant therapy, such as prophylactic cranial irradiation.

Measurement of chemoresistance markers in patients with stage iii non–small cell lung cancer: a novel approach for patient selection

BACKGROUND The long-term survival of patients with stage III non-small cell lung cancer treated with a combination of chemotherapy and radiation is 10% to 20%. Survival could potentially be increased and toxicity limited if one could identify patients most likely to respond to a particular treatment regimen. This project prospectively evaluated a panel of potential immunohistochemical markers of chemoresistance in a population of patients with pathology-confirmed stage III non-small cell lung cancer in order to determine the prognostic value of each marker in relation to response to chemotherapy or survival. METHODS Immunohistochemical staining was performed on histologically positive mediastinal nodal specimens obtained from 59 patients (mean age, 62 years; range, 41 to 79 years) without evidence of distant metastatic disease treated with navelbine-based chemotherapy and external beam radiation therapy between 1996 and 2001. Included were markers for apoptosis (p53, bcl-2), drug efflux/degradation (MDR, GST-pi), growth factors (EGFr, Her2-neu), and mismatch repair (hMLH1, hMSH2). After chemotherapy, patients underwent radiologic evaluation for response measured by standard criteria. RESULTS After a median 41 months of follow-up (range, 17 to 55 months), 43 patients had recurrent disease and 38 of these patients were dead of cancer (median cancer-free survival of 10 months and overall survival of 18 months). Patients who demonstrated a complete or partial response (n = 38) had a significantly improved survival (p = 0.002) compared with those with stable or progressive cancer (n = 21). Multivariable Cox step-wise regression analysis of marker expression associated overexpression of p53 and low expression of hMSH2 with poor treatment response and cancer death. CONCLUSIONS These preliminary data suggest that marker expression may allow the separation of patients into low- and high-risk groups with respect to survival after combined navelbine-based chemotherapy and XRT. This could represent a novel method of selecting patients for a particular treatment regimen if these data are reproduced in a larger prospective trial.

Estrogen and progesterone receptor status determined by the Ventana ES 320 automated immunohistochemical stainer and the CAS 200 image analyzer in 236 early-stage breast carcinomas: Prognostic significance

The quantitation of estrogen and progesterone receptors (ER and PgR) has become the standard of care in the evaluation of patients with primary breast carcinoma. It has been demonstrated that ER and PgR detected by immunohistochemical methods in formalin‐fixed paraffin‐embedded tissue can be quantified by computerized image analysis. In this study, ER and PgR levels were determined by using an automated immunohistochemistry stainer (Ventana ES 320) and an image analyzer (CAS 200) in a series of 236 patients with stage I/II carcinoma of the breast. The degree of correlation of the ER and PgR levels determined by the dextran‐coated charcoal method (DCC) with image analysis quantitation was high (r = 0.75). The agreement between both methods was 77% for ER and 73% for PgR. Hormone receptor levels were correlated with prognosis as determined by overall survival. An ER level of 30 fmol/mg as determined by image analysis was established to stratify the patient population most effectively into favorable and unfavorable prognostic groups (P = 0.003). An ER level of 20 fmol/mg for prognostic stratification reached statistical significance (P = 0.03). The DCC method was not able to stratify the patients into prognostic groups at the traditionally accepted cutpoint of 10 fmol/mg (P = 0.52). We conclude that when used in combination, automated immunohistochemistry and quantitative image analysis offer a favorable alternative to the DCC method in assessment of ER and PgR status in human mammary carcinoma. In addition, quantitative immunocytochemistry techniques may prove superior to the DCC method in specimens in which there is limited tumor volume (including fine‐needle aspirates), stromarich tumors, and early‐stage lesions including intraductal carcinoma.

Neighborhood-level socioeconomic determinants impact outcomes in nonsmall cell lung cancer patients in the Southeastern United States

Studies examining the impact of lower socioeconomic status (SES) on the outcomes of patients with nonsmall cell lung cancer (NSCLC) are inconsistent. The objective of this study was to clearly elucidate the association between SES, education, and clinical outcomes among patients with NSCLC.

Prognostic value of MIB‐1 in advanced ovarian carcinoma as determined using automated immunohistochemistry and quantitative image analysis

The monoclonal antibody MIB‐1 is an immunohistochemical marker reacting most strongly with cells in late S phase, G2, and M portions of the cell cycle. This antibody, reactive in formalin‐fixed, paraffin‐embedded tissue, allows the quantitation of a proliferation index (PI) in both current clinical cases and archival material using a computerized image analyzer (CIA).

Determination of Proliferation Index By MIB‐1 Immunostaining in Early Stage Breast Cancer Using Quantitative Image Analysis

Abstract: Several clinicopathologic variables influence prognosis in breast cancer, including stage, histologic grade, nodal status, and tumor size. Multiple studies have shown an independent value of proliferation index as a prognostic variable for the stratification into favorable and unfavorable groups. The monoclonal antibody MIB‐1 reacts with the same antigen site, not epitope, as recognized by the Ki‐67 antibody. Like Ki‐67, MIB‐1 reacts with cells in the late G1, S, M and G2 phases of the cell cycle, but MIB‐1 has the advantage of reacting with formalin‐fixed, paraffin‐embedded material. The authors investigated the feasibility of using image analysis to quantitate the MIB‐1 antibody staining (proliferation index [PI]) and predict survival in a series of 230 patients with stage I and stage II breast cancer. In a univariate Cox regression model, larger values of MIB‐1 were related to shorter survival times (p < 0.001). Exploratory statistical procedures were used to categorize the patients into good, intermediate, and poor survival groups using the following proliferation indices as cut‐points: <5%, 5–11%, and >11 %, respectively. Higher clinical stage was associated with higher MIB‐1 values and shorter survival (p = 0.01, and p = 0.003, respectively). Tumor size (p = 0.02) and nodal status (p = 0.05) were also associated with higher values of MIB‐1. After adjusting for age, clinical stage, nodal status, and tumor size in a multivariate analysis, MIB‐1 retained its prognostic significance (p < 0.0001) when considered as either a continuous or categorical variable. There were no significant associations between MIB‐1 determined proliferation index and age (p = 0.54), histologic grade (p = 0.69), nuclear grade (p = 0.06) or the presence of vascular invasion (p =.66). There is a strong statistical relationship between cell proliferative activity, as determined by MIB‐1 expression, and survival in early stage breast cancer.

Immunohistochemically Determined Estrogen and Progesterone Receptor Levels: A Comparison of Three Antibodies with the Ligand‐Binding Assay

Abstract: Traditionally, estrogen and progesterone receptor levels have been determined by biochemical ligandbinding assays, but more recently immunohistochemical techniques have become available. They have gained popularity due to their low cost, smaller sample size requirements, and direct visualization capability of reaction location. Several antibody clones are commercially available and antibodies directed against the estrogen receptor (ER) are supplied by Ventana Medical Systems (Tucson, AZ), Abbott Laboratories (Abbott Park, IL), and lmmunotech Westbrook, ME). Antibodies directed against the progesterone receptor (PgR) are supplied by Ventana Medical Systems (Tucson, AZ), lmmunotech (Westbrook, ME), and Becton‐DickinsonKelI Analysis Systems (San Jose, CA). Computer‐assisted image analysis using the CAS ZOOTM (Becton‐DickinsonKIS, San Jose, CA) allows quantitation of immunohistochemically determined receptor levels. Correlation of quantitated immunohistochemical ER levels with values determined by ligand‐binding assay revealed the Ventana antibody to most closely predict the ligand‐binding results (wk = .667). The Ventana anti‐progesterone antibody quantitation most closely correlated with the ligand‐binding results (wk = 435) for determination of PgR. Progesterone receptor level as determined by any of the tested methods did not stratify patients into favorable and unfavorable prognostic groups. Estrogen receptor level as determined by the Ventana antibody was the most predictive of patient outcome but this relationship did not reach statistical significance (p = .09). Most discrepancies between the ligand‐binding assay and the immunohistochemical assays were associated with one of three factors: (a) low volume of neoplastic cells present due either to small sample size or high stromal content, (b) premenopausal status with circulating endogenous estrogens potentially occupying receptor sites, (c) presence of benign breast epithelium resulting in a false‐positive ligand‐binding assay.

Monitoring tumor markers in serial sera predicts disease failure in lung cancer patients following surgery

7040 Background: Patients with early stage non-small cell lung cancer (NSCLC) who undergo resection have a recurrence rate of 50% at 5 years. A contributing factor to such a high recurrence rate is...