Use of FISH Analysis for Diagnosis of Renal Cell Carcinoma Subtypes

Renal cell carcinoma: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up†.

IDENTIFICATION OF TUMOR ENTITIES OF RENAL CELL CARCINOMA USING INTERPHASE FLUORESCENCE IN SITU HYBRIDIZATION

Prognostic Factors of Papillary Renal Cell Carcinoma: Results From a Multi-Institutional Series After Pathological Review

Renal cell carcinoma: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up

Histopathology of Surgically Treated Renal Cell Carcinoma: Survival Differences by Subtype and Stage

Introduction: Renal cell carcinoma (RCC) accounts for 90% of all kidney cancers. It comprises a heterogeneous group of renal tumors with distinct genetic and molecular characteristics including clear cell RCC (ccRCC), papillary RCC (pRCC) and chromophobe RCC (chRCC). The differential diagnosis of RCC subtypes relies on distinct morphology which is not always accurate. Accurate classification of RCC subtypes is critical since each exhibits different clinical behaviour, prognosis and response to therapy. The purpose of this study is to determine whether a limited number of miRNAs can classify RCC subtypes with high accuracy. Experimental Design: We extracted RNA from 90 formalin-fixed paraffin-embedded (FFPE) tissues including 27 clear cell RCC, 29 papillary RCC, 19 chromophobe RCC, 4 unclassified RCC tumors and 11 oncocytomas. We measured the absolute expression of six miRNAs by qRT-PCR. Receiver operator characteristic curves were constructed and the area under the curve (AUC) was calculated to assess diagnostic performance. We also tested miRNA expression by in situ hybridization (ISH) in an independent set of ninety-eight FFPE renal tumors. Results: We developed a two-step miRNA classifier. In the first step, expressions of selected miRNAs were found to discriminate clear cell RCC and papillary RCC from chromophobe RCC and renal oncocytoma. Two miRNAs were able to discriminate clear cell RCC and papillary RCC from chromophobe RCC and oncocytoma. miR-221 was significantly overexpressed in chromophobe RCC and oncocytoma compared to clear cell RCC and papillary RCC (4.49-fold change, p= 6.398e-010) and was able to discriminate between the two groups (AUC: 0.9637, 95% CI: 0.9132 to 1.014, p<0.0001). In the second step, the absolute expression of two miRNAs could distinguish clear cell RCC from papillary RCC (10.4-fold change, p = 1.243e-013). Moreover, an additional two miRNAs could differentiate chromophobe RCC from renal oncocytoma (3.30- fold change, p = 1.751e-006). In situ hybridization revealed that miRNAs display a nuclear staining pattern that was able to distinguish clear cell RCC from papillary RCC (p<0.001) and chromophobe RCC from renal oncocytoma (p =0.009). Conclusion: miRNA expressions were able to distinguish between RCC subtypes and renal oncocytoma. miRNA assessment by in situ hybridization is a clinically useful diagnostic tool that can complement current methods for RCC classification. Citation Format: Ashley Di Meo, Mereet Hanna, Rola Saleeb, Samantha Wala, Adriana Krizova, Manal Gabril, Haiyan Zhai, Maria Pasic, Andrew Evans, Fadi Brimo, George Yousef. A microRNA signature for the classification of renal cell carcinoma subtypes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5447. doi:10.1158/1538-7445.AM2017-5447

Differences in Organ System of Distant Metastasis by Renal Cell Carcinoma Subtype

Diagnosis of uncommon renal epithelial neoplasms: performances of fluorescence in situ hybridization

The clinical staging of renal cortical tumors traditionally has not evaluated the potential effect of histological subtypes on survival. Evidence suggests that conventional clear cell renal cell carcinoma (RCC) and nonconventional clear cell RCC (chromophobe and papillary) have different metastatic potential. Using a large renal tumor database, we examined the effect of tumor histology on the pattern of metastasis and patient survival. All patients with nonmetastatic renal cortical tumors undergoing partial or radical nephrectomy were identified from a renal tumor database between July 1989 and July 2002. Kaplan-Meier and Cox regression tests were used for statistical analysis. Analysis revealed 1057 patients: 794 with conventional clear cell RCC, 157 with papillary RCC, and 106 with chromophobe RCC. Metastasis occurred in 95 conventional clear cell RCC, 9 papillary RCC, and 6 chromophobe RCC with a median follow-up of 34.6, 43.0, and 33.2 months, respectively. Using log-rank analysis, chromophobe and papillary RCC were associated with an improved disease-free survival at 5 years (P =.009 and.015, respectively). Multivariate analysis revealed tumor size, stage, and chromophobe histology as significant variables for disease progression. Renal cortical tumors have distinct histological subtypes with varying degrees of metastatic potential. Conventional clear cell RCC, which comprises two thirds of renal cortical tumors presenting with localized disease, has a less favorable outcome when compared with papillary and chromophobe RCC. Controlling for size and stage, chromophobe, and not papillary, RCC was a significant variable for disease progression compared with conventional clear cell RCC. Knowledge of renal cortical tumor histological subtype is critical for projecting prognosis, tailoring follow-up strategies, and designing clinical trials.

To retrospectively evaluate whether the enhancement patterns of pathologically proved clear cell, papillary, and chromophobe renal cell carcinomas (RCCs) measured on clinical dynamic contrast agent-enhanced magnetic resonance (MR) images permit accurate diagnosis of RCC subtype. This study was Institutional Review Board approved and HIPAA compliant; informed consent was waived. One hundred twelve patients (76 men, 36 women; age range, 25-88 years; mean age, 58.1 years) underwent MR imaging of 113 renal masses (mean diameter, 5.4 cm) with pathologic diagnoses of clear cell (n = 75), papillary (n = 28), or chromophobe (n = 10) RCC. A 1.5-T clinical MR protocol was used before and after (corticomedullary and nephrographic phases) intravenous administration of contrast agent. Region-of-interest measurements within tumor and uninvolved renal cortex were used to calculate percentage signal intensity change and tumor-to-cortex enhancement index. Subtype groups were compared by using linear mixed-effects models. Receiver operating characteristic (ROC) curve analysis was performed for the comparison of clear cell and papillary RCCs. On both the corticomedullary and nephrographic phase images, clear cell RCCs showed greater signal intensity change (205.6% and 247.1%, respectively) than did papillary RCCs (32.1% and 96.6%, respectively) (P < .001). Chromophobe RCCs showed intermediate change (109.9% and 192.5%, respectively). The tumor-to-cortex enhancement indexes at corticomedullary and nephrographic phases were largest for clear cell RCCs (1.4 and 1.2, respectively), smallest for papillary RCCs (0.2 and 0.4, respectively), and intermediate for chromophobe RCCs (0.6 and 0.8, respectively). Signal intensity changes on corticomedullary phase images were the most effective parameter for distinguishing clear cell and papillary RCC (area under ROC curve, 0.99); a threshold value of 84% permitted distinction with 93% sensitivity and 96% specificity. Clear cell, papillary, and chromophobe RCCs demonstrate different patterns of enhancement on two-time point clinical dynamic contrast-enhanced MR images, allowing their differentiation with high sensitivity and specificity.

Focus on kidney cancer

Our objective was to compare cancer-specific survival and to examine associations with outcome among the histologic subtypes of renal cell carcinoma (RCC). We studied 2385 patients whose first surgery between 1970 and 2000 was a radical nephrectomy for sporadic, unilateral RCC. All RCC tumors were classified following the 1997 Union Internationale Contre le Cancer and American Joint Committee on Cancer guidelines. There were 1985 (83.2%) patients with clear cell, 270 (11.3%) with papillary, 102 (4.3%) with chromophobe, 6 (0.3%) with collecting duct, 5 (0.3%) with purely sarcomatoid RCC and no underlying histologic subtype, and 17 (0.7%) with RCC, not otherwise specified. Cancer-specific survival rates at 5 years for patients with clear cell, papillary, and chromophobe RCC were 68.9%, 87.4%, and 86.7%, respectively. Patients with clear cell RCC had a poorer prognosis compared with patients with papillary and chromophobe RCC (p <0.001). This difference in outcome was observed even after stratifying by 1997 tumor stage and nuclear grade. There was no significant difference in cancer-specific survival between patients with papillary and chromophobe RCC (p = 0.918). The 1997 TNM stage, tumor size, presence of a sarcomatoid component, and nuclear grade were significantly associated with death from clear cell, papillary, and chromophobe RCC. Histologic tumor necrosis was significantly associated with death from clear cell and chromophobe RCC, but not with death from papillary RCC. Our results demonstrate that there are significant differences in outcome and associations with outcome for the different histologic subtypes of RCC, highlighting the need for accurate subtyping.

Objective To analyze the clinicopathological features and prognostic factors of common subtypes of non-transparent renal cell carcinoma. Methods Retrospective analysis of 115 patients with pathologically confirmed non-transparent renal cell carcinoma from January 2003 to December 2017, including 67 males and 48 females, with a male to female ratio of 1.4∶1. The average age is (51.2±13.4) years old. 71 cases were asymptomatic renal cancer, 44 cases had clinical symptoms, including 10 cases of gross hematuria, 28 cases of low back pain, 4 cases of hematuria with low back pain, and 2 cases of abdominal mass. There were 49 open surgery and 66 laparoscopic surgery. 58 patients underwent radical nephrectomy and 57 underwent partial nephrectomy. Of the 115 patients, 17 (14.9%) had abnormal hemoglobin (Hb), 22 (19.1%) had abnormal platelet (PLT) count, 18 (15.7%) had abnormal alkaline phosphatase, and abnormal lactate dehydrogenase 16 cases (13.9%). The Kaplan-Meier survival analysis method was used to calculate the survival rate of patients, and the Cox proportional regression risk model was used to analyze the prognostic factors. Results The postoperative pathological stage was 57 cases in T1a stage, 38 cases in T1b stage, 12 cases in T2a stage, 8 cases in T2b stage, 2 cases of regional lymph node positive, and 113 cases negative; no distant metastasis. Pathological types: 42 cases of renal chromophobe cell carcinoma, 37 cases of papillary renal cell carcinoma type Ⅰ, 36 cases of type Ⅱ. The average follow-up time was 38.6 months, and the rate of loss of follow-up was 3.5% (4/115). The 1, 3, and 5 year overall survival rates of 115 patients with common subtypes of non-transparent renal cell carcinoma were 99.1%, 95.8%, and 81.1%, respectively. Multivariate Cox regression analysis found that the pathological type (OR=4.625, P=0.014), four indicators ≥3 abnormalities (OR=30.853, P=0.024), lymph node metastasis (OR=35.663, P=0.006) were the group. An independent factor in the survival time of patients with common subtypes of non-transparent renal cell carcinoma. Conclusions Compared with papillary renal cell carcinoma type Ⅰ and renal chromophobe cell carcinoma, papillary renal cell carcinoma type Ⅱ has a higher degree of malignancy and a poor prognosis. The pathological types of the common subtypes of non-transparent renal cell carcinoma, four indicators (Hb, PLT count, alkaline phosphatase, and lactate dehydrogenase)≥3 abnormalities and lymph node metastasis are independent prognostic factors for overall survival. Key words: Non-transparent renal cell carcinoma; Renal chromophobe cell carcinoma; Papillary renal cell carcinoma; Prognosis

Objective To analyze the value of multi-slice spiral CT (SCT) scan in staging and subtyping of renal cell carcinoma (RCC). Methods The preoperative kidney SCT data and postoperative pathology results of 64 patients with RCC were retrospectively analyzed. The patients′ ages ranged from 33-78 years (average 54 years). There were 44 males and 20 females in the study group. According to the CUA Guidelines, the staging and subtyping of RCC were performed through the combined information of preoperative SCT attenuation in unenhanced, corticomedullary phase and enhancement pattern. The results were compared with the postoperative histopathological results. Results The SCT results showed 38 cases were clear cell RCC, 14 cases were papillary RCC and 12 cases were chromophobic cell RCC. Histopathological results showed that 40 cases were clear cell RCC, 16 cases were papillary RCC and 8 cases were chromophobic cell RCC. According to the standard of 40 HU of CT attenuation value, the sensitivity, specificity and accuracy were 75%, 79% and 78% for diagnosis of papillary RCC in the unenhanced phase. The sensitivity, specificity and accuracy by the standard of 90 HU of CT attenuation value was 90%, 88% and 89% for diagnosis of clear cell RCC in the corticomedullary phase. In chromophobic RCC, homogeneous enhancement was more common than in papillary RCC and clear cell RCC. There was no significant difference of staging and subtyping of RCC between SCT and pathological results (P>0.05). The accuracy of SCT in staging and subtyping of RCC was 88% in staging, and 89% in subtyping. Conclusions SCT is a useful preoperative tool to stage and subtype RCC Key words: Kidney neoplasms; Carcinoma; Tomography, X-ray computed; Neoplasm staging

We investigated the role of fluorescence in situ hybridization (FISH) in the diagnosis of primary renal neoplasms and lesions suspicious for metastatic renal cell carcinoma. Consecutive fine-needle aspiration biopsies (FNAB) of 39 renal masses and 41 metastatic tumours suspicious for renal cell origin were assessed with an immunohistochemical panel for CK7, RCC antigen, CD10, AMACR, PAX8, vimentin, and CD117. In addition, FISH was performed using probes for chromosomes 1p, 3p, 7, 17, X, and Y. A total of 31 of 39 primary renal masses and 33 of 41 metastatic tumors suspicious for renal origin demonstrated typical cytological and immunohistochemical (IHC) features of subtypes of renal neoplasms (40 clear cell renal cell carcinoma (RCC), 20 papillary RCC, and 4 renal oncocytomas). FISH analysis of 15 randomly selected cases each of primary and metastatic lesions revealed chromosomal abnormalities consistent with the diagnosis in 73% of these cases. Of 8 primary renal masses demonstrating atypical microscopic features and noncontributory IHC profiles, FISH was helpful in subtyping 5 (62%) of these lesions (2 clear cell RCC, 1 solid variant of oncocytic papillary RCC, 1 mixed clear cell and papillary RCC, and 1 chromophobe RCC with papillary architecture). Of 8 metastatic tumors clinically suspicious for renal cell origin and supportive, but nondiagnostic IHC, FISH revealed supportive chromosomal changes in 6 (75%) cases. In conclusion FISH analysis on FNAB material, even with limited tissue, may be contributory to the diagnosis and subtyping of RCC in diagnostically challenging biopsies.