Indications and results of diagnostic biopsy in pediatric renal tumors: A retrospective analysis of 317 patients with critical review of SIOP guidelines.

OBJECTIVES: According to the Renal Tumor Study Group (RTSG) of the International Society of Paediatric Oncology (SIOP), diagnostic biopsy of renal tumors prior to neoadjuvant chemotherapy is not mandatory unless the presentation is atypical for a Wilms tumor (WT). This study addresses the relevance of this strategy as well as the accuracy and safety of image-guided needle biopsy. METHODS: Clinical, radiological, and pathological data from 317 children (141 males/176 females, mean age: 4 years, range, 0-17.6) consecutively treated in one SIOP-affiliated institution were retrospectively analyzed. RESULTS: Presumptive chemotherapy for WT was decided for 182 patients (57% of the cohort), 24 (8%) were operated upfront, and 111 (35%) were biopsied at diagnosis. A non-WT was confirmed after surgery in 5/182 (3%), 11/24 (46%), and 28/111 (25%), respectively. Age at diagnosis was the most commonly (46%) used criterion to go for biopsy but a nine-year threshold should be retrospectively considered more relevant. Tumor volumes of clear cell sarcoma of the kidney and WT were significantly higher than those of other tumors (P = 0.002). The agreement between core-needle biopsy (CNB) and final histology was 99%. No significant morbidity was associated with CNB. CONCLUSION: The use of SIOP criteria to identify patients eligible for presumptive WT neoadjuvant chemotherapy or upfront surgery avoided biopsy in 65% of children and led to a 97% rate of appropriate preoperative chemotherapy. Image-guided CNB is a safe and accurate diagnostic procedure. The relevance of SIOP biopsy criteria might be improved by using an older age threshold (9 years instead of 6 years) and by adding initial tumor volume.

Prognostic value of tumor mutational burden in patients with oral cavity squamous cell carcinoma treated with upfront surgery.

BACKGROUND: Oral cavity is the most prevalent site of head and neck squamous cell carcinomas (HNSCCs). Most often diagnosed at a locally advanced stage, treatment is multimodal with surgery as the cornerstone. The aim of this study was to explore the molecular landscape of a homogenous cohort of oral cavity squamous cell carcinomas (OCSCCs), and to assess the prognostic value of tumor mutational burden (TMB), along with classical molecular and clinical parameters. PATIENTS AND METHODS: One hundred and fifty-one consecutive patients with OCSCC treated with upfront surgery at the Institut Curie were analyzed. Sequencing of tumor DNA from frozen specimens was carried out using an in-house targeted next-generation sequencing panel (571 genes). The impact of molecular alterations and TMB on disease-free survival (DFS) and overall survival (OS) was evaluated in univariate and multivariate analyses. RESULTS: Pathological tumor stage, extranodal spread, vascular emboli, and perineural invasion were associated with both DFS and OS. TP53 was the most mutated gene (71%). Other frequent molecular alterations included the TERT promoter (50%), CDKN2A (25%), FAT1 (17%), PIK3CA (14%), and NOTCH1 (15%) genes. Transforming growth factor-ß pathway alterations (4%) were associated with poor OS (P = 0.01) and DFS (P = 0.02) in univariate and multivariate analyses. High TMB was associated with prolonged OS (P = 0.01 and P = 0.02, in the highest 10% and 20% TMB values, respectively), but not with DFS. Correlation of TMB with OS remained significant in multivariate analysis (P = 0.01 and P = 0.005 in the highest 10% and 20% TMB values, respectively). Pathological tumor stage combined with high TMB was associated with good prognosis. CONCLUSION: Our results suggest that a high TMB is associated with a favorable prognosis in patients with OCSCC treated with upfront surgery.

Comprehensive genomic profiling of head and neck squamous cell carcinoma reveals FGFR1 amplifications and tumour genomic alterations burden as prognostic biomarkers of survival.

BACKGROUND: We aimed at identifying deleterious genomic alterations from untreated head and neck squamous cell carcinoma (HNSCC) patients, and assessing their prognostic value. PATIENTS AND METHODS: We retrieved 122 HNSCC patients who underwent primary surgery. Targeted NGS was used to analyse a panel of 100 genes selected among the most frequently altered genes in HNSCC and potential therapeutic targets. We selected only deleterious (activating or inactivating) single nucleotide variations, and copy number variations for analysis. Univariate and multivariate analyses were performed to assess the prognostic value of altered genes. RESULTS: A median of 2 (range: 0-10) genomic alterations per sample was observed. Most frequently altered genes involved the cell cycle pathway (TP53 [60%], CCND1 [30%], CDKN2A [25%]), the PI3K/AKT/MTOR pathway (PIK3CA [12%]), tyrosine kinase receptors (EGFR [9%], FGFR1 [5%]) and cell differentiation (FAT1 [7%], NOTCH1 [4%]). TP53 mutations (p = 0.003), CCND1 amplifications (p = 0.04), CDKN2A alterations (p = 0.02) and FGFR1 amplifications (p = 0.003), correlated with shorter overall survival (OS). The number of genomic alterations was significantly higher in the HPV-negative population (p = 0.029) and correlated with a shorter OS (p < 0.0001). Only TP53 mutation and FGFR1 amplification status remained statistically significant in the multivariate analysis. CONCLUSION: These results suggest that genomic alterations involving the cell cycle (TP53, CCND1, CDKN2A), as well as FGFR1 amplifications and tumour genomic alterations burden are prognostic biomarkers and might be therapeutic targets for patients with HNSCC.

Quantitative mapping of 4'-iododeoxyrubicin in metastatic squamous cellcarcinoma by secondary ion mass spectrometry (SIMS) microscopy.

Secondary ion mass spectrometry microscopy enables quantitative mapping of chemical elements in tissue sections. It was used for the detection of 127I contained in 4'-iododeoxyrubicin (IDX). Metastatic cutaneous squamous cell carcinoma from 7 patients participating in a phase I study (IDX dose, 80 mg/m2, 10-min i.v. infusion) were biopsied 10 min after drug administration and compared to 3 controls who did not receive any treatment (one squamous cell carcinoma and 2 gastric carcinomas). Biopsy specimens were fixed and embedded in methacrylate resin. Then, serial semithin sections (3 microns) were analyzed simultaneously with ionic and optimal microscopes in order to identify the histological structures given by the 31P distribution in which 127I in IDX was mapped. The iodine signal was undetected in controls and found mainly in the nuclei of tumor cells of the treated patients. Its concentration, measured in at least 30 nuclei of each specimen, was undetectable in 8% of the nuclei and 91% of them were within 1 and 16 ng/mg. The mean concentration of each specimen ranged from 5 to 23 ng/mg. This study demonstrates the capacity of ion microscopy to localize a cytotoxic drug (IDX) in a human biopsy specimen without the need for radioactive labeling and enables the evaluation of drug penetration in cancer cells which is critical for its activity.

Alterations of the DNA repair gene OGG1 in human clear cell carcinomas of the kidney.

The OGG1 gene, which codes for a DNA repair protein with antimutator activity, is located on chromosome 3p25, a frequent site of allelic deletions in many types of human tumors, including renal clear cell cancers. We present the analysis of 99 renal tumors for alterations in the OGG1 gene to determine its association with tumorigenesis. Loss of heterozygosity in the 3p25 region was found for 85% of the informative cases. We detected somatic missense mutations of the OGG1 gene in 4 of the 99 tumor samples. Biochemical analysis of the mutant proteins revealed that a substitution at codon 46 impairs the enzymatic activity. We also describe the occurrence of several polymorphisms as well as aberrantly spliced OGG1 transcripts.

Biological and clinical significance of concurrent p53 gene alterations, MDR1 gene expression, and S-phase fraction analyses in breast cancer patients treated with primary chemotherapy or radiotherapy.

We investigated the interrelationship between p53 gene alterations, MDR1 gene expression, and S-phase fraction (SPF) in breast carcinomas treated primarily with chemotherapy or radiotherapy and correlated the results with patient outcome to determine the potential clinical significance of these factors. In a consecutive series of 64 fine-needle samplings of breast cancer patients who underwent either neoadjuvant chemotherapy (n = 53) or radiotherapy (n = 11), p53 (exons 5-9) gene alterations by denaturating gradient gel electrophoresis and subsequent direct sequencing, MDR1 gene expression by semiquantitative reverse transcription-PCR, and SPF by DNA flow cytometry were determined. Our results show that p53 mutations (n = 20) were significantly associated (P = 0.01) with high SPF but not with de novo MDR1 gene expression. Most patients with wild-type p53 tumors were found to be resistant to neoadjuvant chemotherapy. No correlation was observed between p53 mutations and the induction of MDR1 gene expression during treatment. Although a significant correlation between shorter distant disease-free survival and high (>/=5%) SPF (P = 0.016) was found, no correlation between distant disease-free survival and p53 status or intrinsic MDR1 gene expression was found. Poor overall survival was observed in patients with tumors with high SPF (P < 0.0004) or lacking MDR1 gene expression (P = 0.03) before treatment, but not with p53 alterations. These data suggest that SPF remains the most relevant biological factor for breast cancer patients treated by primary chemotherapy or radiotherapy and that p53 and MDR1 status may identify a small subset of patients that may resist therapy or pursue an aggressive course.

[Pseudotumoral soft tissue masses in children and adolescents]. / Masses des tissus mous d'allure tumorale de l'enfant et de l'adolescent.

INTRODUCTION: Pseudotumoral soft tissue masses in children and adolescents are a frequent reason for consultation and a diagnostic dilemma. Soft tissue malignancies are relatively uncommon, unlike the large number of benign lesions that may be seen in the superficial tissue and that can be diagnosed with clinical characteristics. MATERIALS AND METHODS: This retrospective study concerns 161 children and adolescents less than 20 years old, referred for a soft tissue mass between 2007 and 2011. It describes their epidemiology, clinical characteristics, and course of care to validate a diagnostic strategy for such masses. RESULTS: Final diagnoses were malignant tumors (44%), benign tumors (32%), and pseudotumoral lesions (24%). Clinical features were similar between these three groups except for age and tumor location, with more benign thoracic masses in younger children. Clinical and radiological association led to an accurate diagnosis for 50% of benign masses and with cytological analysis contribution in 79% of benign tumors and 86% of pseudotumoral lesions. Malignant tumors were suspected in only 39% of cases with radiological exams and in 89% after fine-needle aspiration, an essential additional diagnostic tool. Final diagnoses were formally established through simple standard clinical and radiological evaluation in 19 patients (11.8%; benign tumors, seven patients; malformations, eight patients; post-traumatic lesions, two patients; infection and inflammation, one patient each); ultrasound exam in five patients (3.1%; hemangioendotheliomas, two patients, fascial dehiscence, hemangioma, and vascular malformation, one patient each); MRI in four patients (2.5%; three vascular malformations and one lipoma); CT in two cases (1.2%; vascular malformation and myositis ossificans), and radiological examinations associated with cell aspiration in 15 cases (9.3%; ten benign tumors and five malignant tumors). CONCLUSIONS: A multidisciplinary approach should be requested from oncological, radiological, and pathologic experts to optimize soft tissue mass management as soon as initial investigations start. The authors advise a diagnostic strategy for children with pseudotumoral soft tissue masses.

Serological and immunohistochemical detection of a 65-kDa oncofetal protein in breast cancer.

Sera from 132 breast cancer patients and 112 healthy female controls were tested with a double-antibody sandwich ELISA using two different monoclonal antibodies against the 65-kDa oncofetal protein, termed p65. Of cancer sera, 90.2% were positive for p65. The average level of p65 was 466.5 +/- 243.8 ng/ml (mean +/- S.D.) in carcinomas and 37.4 +/- 29.5 ng/ml (mean +/- S.D.) in controls (P < 0.0005). A selected group (n = 15) of these 132 patients were needle-biopsied and assessed immunohistochemically using monoclonal antibodies against p65. Nucleocytoplasmic expression was found in 12 patients (80%) using monoclonal antibodies. Expressions of p65 were concordant in 13 (86%) cases between serum and tumour tissues, but did not correlate with tumour DNA ploidy, histological grade or hormone receptors levels. Sera were also tested for CA 15-3 with the average value in cancer serum being 132.4 +/- 14.0 U/ml; there was no significant concordance between the two markers. Thus, p65 may be a potential serum and/or immunohistochemical marker for breast carcinoma.

In vitro radiation-induced apoptosis and early response to low-dose radiotherapy in non-Hodgkin's lymphomas.

PURPOSE: Prospective investigation of spontaneous and in vitro radiation-induced apoptosis to predict early response to palliative radiotherapy in patients with non-Hodgkin's lymphomas. PATIENTS AND METHODS: Fine-needle sampling was performed in 28 tumor sites (26 patients) and yielded adequate cell numbers in 27 cases. Apoptotic cells were counted by fluorescence microscopy immediately after sampling and after 24-h culture (spontaneous apoptosis) and 24 h after 2- and 10-Gy in vitro irradiation (radiation-induced apoptosis). Early response to low-dose in vivo radiotherapy (mostly 4 Gy in two fractions over 3 days) was evaluated 15 days after treatment. RESULTS: The tumor response rates at 15 days were 11 (39%) complete responses, nine (32%) responses of greater than 50% reduction in volume, six (21%) responses of less than 50% reduction in volume and two (7%) cases of no response. Tumors achieving complete or major response after in vivo irradiation had higher percentages of apoptotic cells after in vitro irradiation, while no significant differences in terms of spontaneous apoptosis were observed between responders and non-responders. CONCLUSION: Spontaneous and in vitro radiation-induced apoptosis can be easily and quickly assessed on cells obtained by fine-needle sampling of non-Hodgkin's lymphoma lesions. The present results suggest that in vitro radiation-induced apoptosis could be used as a predictive assay of early response to low-dose in vivo irradiation in patients with non-Hodgkin's lymphomas.

Immunoenzymatic (EIA) assays of estrogen and progesterone receptors in fine-needle and surgical samples in breast cancer patients.

Both preoperative cytological (FNS) and tumorectomy surgical specimens of the same tumor were obtained from 89 patients with breast cancer. Estrogen and progesterone receptors were assayed on cytosolic extracts by enzyme immunoassay (EIA), in 89 patients for estrogen and in 68 patients for progesterone. Both concentrations were highly correlated between the FNS and the corresponding surgical sample, but FNS values yielded 2 to 3 times higher. With cut-off values of 250 fmol/mg DNA, corresponding to 15 fmol/mg cytosol protein for surgical samples and 500 fmol/mg DNA for FNS, the receptor status was concordant between the types of samples in 84 of 89 (94%) and 59 of 68 (86.8%) patients for estrogen and progesterone, respectively. Sixty-seven carcinomas were estrogen positive, 17 estrogen negative, 38 progesterone positive, and 21 were progesterone negative in both samples. The authors conclude that estrogen and progesterone EIAs can be performed on FNS provided by properly trained pathologists, with a control of their cellularity by DNA assay.

Analysis of correlation between mitotic index, MIB1 score and S-phase fraction as proliferation markers in invasive breast carcinoma. Methodological aspects and prognostic value in a series of 257 cases.

BACKGROUND: The study was designed in order to evaluate the degree of correlation of mitotic index (MI), Ki67 (MIB1) score and S-phase fraction (SPF) as markers of cell proliferation and prognosis in breast cancer. MATERIALS AND METHODS: The series analysed corresponded to 257 consecutive invasive breast carcinoma, treated at the Institut Curie, France, in 1995. Nottingham histological grade and MIB1 semiquantitative and quantitative score were assessed on histological sections, whereas SPF was calculated using flow cytometry analysis of fine-needle aspiration products. Proliferation indices were compared to pathological data and to overall survival (OS) and disease-free survival (DFS) (minimum follow-up: 72 months). RESULTS: The median values for the proliferation markers were 9/10 HPF for MI, 32.4% for MIB1 and 3.7% for SPF. A high rate of correlation (r=0.96; p<0.001) was observed between semi-quantitative and quantitative MIBI evaluation. A positive correlation was found between the three markers (r ranging from 0.54 to 0.61;p<0.001). Univariate analysis of markers associated to disease outcome showed that MIB1, axillary node status (N) and progesterone receptor (PR) status were significantly associated with OS and that MIB1 and SPF were associated with DFS, together with node and hormone receptor status. In multivariate analysis, when proliferation markers were adjusted on the N and PR status, only MIB1 retained a prognostic value for OS (RR= 1.83) [1.00;3.35] and SPF for DFS (RR= 1.58) [1.02-2.44] (p=0.04). CONCLUSION: A good level of correlation was observed between the values of the three markers of tumour cell proliferation analysed. In this series of invasive breast cancers, MIB1 immunostaining was found to be a prognostic marker of both OS and DFS. The median (32.4%) was a valuable cut-off value for prognostic assessment. Semi-quantitative and quantitative evaluations provided very similar values. MIB1 can thus be considered as a reliable prognostic maker, usable in small size tissue specimens which are inappropriate for MI or SPF analysis. The impact of MIB1 compared to that of the other proliferative markers will be further assessed in a subgroup of T1N0M0 for which the prognostic assessment is of major interest.

So-called branchiogenic carcinoma is actually cystic metastases in the neck from a tonsillar primary.

In 1974, we reported on six patients with squamous cell carcinoma arising in the tonsillar area who presented with a characteristic form of cystic nodal metastasis. This study reports on an additional 21 patients with this entity. Treatment of these patients included removal of the cystic metastasis with or without radical neck dissection and irradiation of Waldeyer's ring. Two patients have died of unrelated disease and two were lost to follow-up, but the remainder are alive without disease 2 months to 12 years following initial diagnosis. We believe that cystic metastases from tonsillar squamous cell carcinomas have often been mistaken for either primary squamous cell carcinomas of branchiogenic origin or with branchial cleft cysts. The distinctive histological features of cystic metastases reviewed after its correct identification can lead to the discovery of an unsuspected primary lesion and result in specific treatment options.

Basaloid squamous carcinoma of the head and neck. Immunohistochemical comparison with adenoid cystic carcinoma and squamous cell carcinoma.

Basaloid squamous carcinoma (BSC) is a rare distinct variant of squamous cell carcinoma of the head and neck region. This entity may commonly pose diagnostic difficulties, especially on small biopsy material. We report the clinicopathological characteristics of 12 new cases and compare their immunohistochemical features with those of solid adenoid cystic carcinomas and conventional squamous cell carcinomas. Our results show that neoplastic BSCs and squamous cell carcinomas do not react to vimentin and S100 protein, while adenoid cystic carcinomas manifest both. The BSCs, however, are S100 protein-positive in intratumoral dendritic Langerhans' cells that are lacking in squamous cell and adenoid cystic carcinomas. Our findings indicate that the immunohistochemical differences between BSC and adenoid cystic carcinoma could assist in their differential diagnosis.

Undifferentiated carcinoma of nasopharyngeal type of tonsil.

A review of 2262 squamous cell carcinomas of the tonsillar region seen at the Institut Gustave-Roussy, Villejuif, France, from 1970 to 1986 showed 1837 well- and poorly differentiated squamous cell carcinomas and 425 undifferentiated squamous cell carcinomas. Eighteen patients with undifferentiated squamous cell carcinomas presented histologic characteristics of undifferentiated carcinomas of nasopharyngeal type. Radiosensibility and radiocurability (complete sterilization with 70 Gy administered) was found in this group with an excellent long-term control of local disease (77% at 10 years actuarial). Epstein-Barr virus-related serologic tests were performed for seven patients. Four of them presented the serologic anti-Epstein-Barr virus titer patterns, generally associated with undifferentiated carcinomas of nasopharyngeal type (1280 to 5120 for viral capsid antigen/IgG and 40 to 320 for viral capsid antigen/IgA). These observations confirm that undifferentiated carcinomas of the nasopharyngeal type may arise outside the nasopharynx.

Fine-needle sampling of salivary gland lesions. VII. Cytology and histology correlation of five cases of epithelial-myoepithelial carcinoma.

Fine-needle sampling (FNS) of five cases of epithelial-myoepithelial carcinoma, three primary tumors, and two local recurrences, was performed preoperatively in five patients. Cytologic diagnoses of malignancy were established in all tumors (three were reported as adenoid cystic carcinoma, two as adenocarcinoma not otherwise specified). Material for cytologic evaluation was satisfactory in all cases. Adenoid cystic carcinoma, polymorphous low-grade adenocarcinoma, papillary cystadenocarcinoma and cellular type of pleomorphic adenoma are the main differential diagnoses.

Salivary carcinomas with papillae: cytology and histology analysis of polymorphous low-grade adenocarcinoma and papillary cystadenocarcinoma.

We retrospectively compared fine-needle samplings (FNS) from two recently individualized low-grade papillary salivary carcinomas, polymorphous low-grade adenocarcinoma (PLGAC) and papillary cystadenocarcinoma (PCAC), to define the cytologic criteria allowing the correct diagnosis. Twelve PLGACs in 10 patients and 5 PCACs in 4 patients were investigated by FNS preoperatively. In both entities, smears contained variable proportions of malignant cells, occasionally arranged in pseudopapillary formations. In contrast to PCAC, PLGAC showed stromal fragments and hyaline globules resembling pleomorphic adenoma or adenoid cystic carcinoma. Cytologic diagnoses of malignancy were established in 10 (83.3%) PLGACs and in 4 (80%) PCACs. Two (16.7%) PLGACs were misdiagnosed as pleomorphic adenoma, and 1 (20%) PCAC as a salivary cyst.

Fine-needle sampling of salivary gland lesions. III. Cytologic and histologic correlation of 75 cases of adenoid cystic carcinoma: review and experience at the Institut Curie with emphasis on cytologic pitfalls.

Fine-needle sampling (FNS) of 75 adenoid cystic carcinomas, including 44 primary tumors, 18 local recurrences, 10 lymph node, and 3 distant metastases, was performed in 66 patients. Concordant cytologic diagnoses were established in 68 tumors (90.7%), whereas 4 (5.4%) were classified as malignant (adenocarcinoma), 1 (1.3%) as suspicious, and 1 (1.3%) as pleomorphic adenoma. The material was insufficient for cytologic evaluation in 1 (1.3%) tumor. Adenoid cystic carcinoma is, in our opinion, easy to diagnose using the FNS technique.

Fine-needle sampling of salivary gland lesions. VI. Cytological review of 44 cases of primary salivary squamous-cell carcinoma with histological correlation.

Fine-needle sampling (FNS) of 44 salivary squamous cell carcinomas, including 34 primary tumors arising from parotid gland, 6 local recurrences, and 4 lymph node cervical metastases, was performed in 44 patients. Cytologic diagnoses were concordant with histological diagnoses in 36 (81.8%) cases, whereas 5 (11.4%) cases were classified as malignant (carcinoma or adenocarcinoma), 1 (2.3%) case as suspicious, and 2 (4.5%) cases were false-negative (necrosis). No samples were unsatisfactory for cytologic evaluation.

Fine-needle sampling of salivary gland lesions. I. Cytology and histology correlation of 412 cases of pleomorphic adenoma.

Fine-needle sampling (FNS) of 412 pleomorphic adenomas, corresponding to 378 primary tumors and 34 local recurrences, was performed preoperatively in 378 patients. Concordant cytologic diagnoses were established in 376 tumors (91.3%), whereas seven (1.7%) were only classified as benign. Sixteen (3.9%) tumors were considered to be suspicious and two (0.5%) were diagnosed as adenoid cystic carcinomas. Insufficient material for cytologic evaluation was found in 11 (2.6%) tumors. Pleomorphic adenomas were retrospectively classified according to the predominant histological differentiation: chondromyxoid, cellular, myoepithelial, or metaplastic. FNS performances were higher in chondromyxoid than in other types, with 95.2% concordant and 1.4% suspicious/ false-positive, vs. 81.8% concordant, and 11.5% suspicious/false-positive cytodiagnoses, respectively.