How to read a next-generation sequencing report-what oncologists need to know.

Next-generation sequencing (NGS) of tumor cell-derived DNA/RNA to screen for targetable genomic alterations is now widely available and has become part of routine practice in oncology. NGS testing strategies depend on cancer type, disease stage and the impact of results on treatment selection. The European Society for Medical Oncology (ESMO) has recently published recommendations for the use of NGS in patients with advanced cancer. We complement the ESMO recommendations with a practical review of how oncologists should read and interpret NGS reports. A concise and straightforward NGS report contains details of the tumor sample, the technology used and highlights not only the most important and potentially actionable results, but also other pathogenic alterations detected. Variants of unknown significance should also be listed. Interpretation of NGS reports should be a joint effort between molecular pathologists, tumor biologists and clinicians. Rather than relying and acting on the information provided by the NGS report, oncologists need to obtain a basic level of understanding to read and interpret NGS results. Comprehensive annotated databases are available for clinicians to review the information detailed in the NGS report. Molecular tumor boards do not only stimulate debate and exchange, but may also help to interpret challenging reports and to ensure continuing medical education.

[Leiomyomatosis of the colon: case report and literature review]. / Leiomyomatose des Rektums: Fallbericht und Literaturübersicht.

We present the findings of a 67 year old male patient with an intestinal leiomyomatosis localized in the rectum. To our knowledge, this is the fifth case of intestinal leiomyomatosis reported so far. The most characteristic findings of this rare disease include a cuff-like tumorous proliferation of smooth muscle within the bowel wall which may extend into extramural tissue and result in a stenosis of a longer bowel segment. Because of severe obstructive symptoms over 5 years, the patient had to undergo surgery with resection of the rectum. The histological examination revealed a morphology and immunophenotype comparable to usual leiomyomas with the exception of hyalinosis-like changes in the blood vessels, apparently a special feature of leiomyomatosis. A novel finding in our case was the occurrence of skeinoid fibers which have so far only been reported in gastrointestinal stromal tumors.

[123I-Metaiodobenzylguanidine- (MIBG-) scintigraphy: paradoxical positivity in an oncocytic adrenocortical carcinoma]. / 123I-Methyljodbenzylguanidin- (MIBG-) Szintigraphie: Paradoxe Nuklidspeicherung eines onkozytären Nebennierenrindenkarzinoms.

(123)I-metaiodobenzylguanidine (MIBG), a radio-labeled catecholamine analogue, is used for the imaging of pheochromocytoma based on the selective uptake of MIBG by chromaffin tissues. MIBG scintigraphy displays high sensitivity (90%) and specificity (close to 100%). In contrast, the false-positive uptake of MIBG by adrenal cortical carcinoma is rare. Here, we report a metastatic oncocytic adrenal cortical carcinoma with MIBG uptake used for therapeutic purposes.

The use of ultrasound and computed tomography for the diagnosis of a squamous cell carcinoma of the oesophago-cardial region of the stomach in a rhesus monkey.

A 5-year-old female rhesus macaque (Macaca mulatta) suddenly began suffering from anorexia, dysphagia, vomiting, diarrhoea, and anaemia. Clinical examination and conventional radiography were uneventful. Additionally an ultrasound (US) and computed tomography (CT) were performed which revealed a large tumorous mass in the upper abdomen and a lung metastasis. Using sonographic guidance, a biopsy of the abdominal masse was taken. Histopathological analysis revealed the diagnosis of a squamous cell carcinoma. At autopsy, an advanced gastric carcinoma, which originated from the cardia, was found with infiltration of the retroperitoneum, and metastatic involvement of the mesenterial lymph nodes as well as metastasis in the lung parenchyma. This case illustrates the usefulness of modern non-invasive imaging techniques, including US and CT, in enabling a quick and accurate diagnosis in laboratory animals.

[Diagnostic relevance of multinucleated giant cells in papillary thyroid cancer. A cytological and histological study]. / Diagnostische Relevanz mehrkerniger Riesenzellen beim papillären SchilddrüsenkarzinomEine zytologische und histologische Untersuchung.

Cytological smears of 17 papillary carcinomas (PC) of the thyroid as well as histological slides of 58 PC and 50 follicular adenomas (FA) and 50 follicular carcinomas (FC) were reviewed to assess the presence of intrafollicularly located multinucleated giant cells. In accordance with the data published, such giant cells were found in 70% of PC but in only 8% of FA and FC. The presence of giant cells, which probably represents a foreign body reaction to, in case of PC, physicochemically altered colloid, is a useful additional criteria for the cytological and histological diagnosis of PC.

[IUD-associated pseudoactinomycotic gradiate granules (Spendore-Hoeppli phenomenon) in uterine curettage]. / Pseudoaktinomykotische Granula (Splendore-Hoeppli-Phänomen) im Korpusabradat bei IUP.

Pseudoactinomycotic radiate granules, also known as the Splendore-Hoeppli phenomenon represent a response to organic and in-organic foreign substances and can be found at various locations in the body. They are frequently observed in uterine curettages, where they can be misinterpreted as actinomycotic sulfur granules. Here we report a case with Splendore-Hoeppli phenomenon in an intrauterine device (IUD)-associated genital actinomycosis. The morphological features of this phenomenon are presented using special staining methods and electron microscopy and the literature is reviewed.

Distribution of HPV 53, HPV 73 and CP8304 in genital epithelial lesions with different grades of dysplasia.

To characterize the risk of malignant progression of cervical epithelial lesions associated with human papillomavirus (HPV) types of yet unknown oncogenic potential the prevalences of these HPVs in different cervical epithelial lesions of 809 patients were determined. HPV types 53, 73, and CP8304 were detected in genital specimens of 16, 22, and 12 of the patients, respectively. The ratio of prevalence in high grade dysplastic lesions or cancers and low grade dysplastic lesions or normal specimens was calculated and compared to corresponding values of well known high-risk (HR) and low-risk (LR) HPVs. For HPV 6, 11, 16, 18, 35, and 73 a ratio of 0.1, 0.2, 5.9, 6.5, 2.5, and 2.4, respectively, was calculated. The ratios of HPV53 and CP8304 were less than 1. Moreover, in contrast to HPV73, these viruses have never been detected in cancer specimens. Thus, HPV53 and CP8304 infections are probably not associated with a high risk of carcinogenesis, while HPV73 could be another HR-HPV type.

[Morphology, DNA cytophotometry and prognosis in gastrointestinal neuroendocrine tumours (carcinoids). A clinico-pathological study of 95 patients]. / Morphologie, DNA-Zytophotometrie und Prognose gastrointestinaler neuroendokriner Tumoren (Karzinoide). Eine klinisch-pathologische Untersuchung an 95 Patienten.

A total of 123 manifestations (97 primary tumours and 26 metastases) of neuroendocrine tumours of the gastrointestinal tract observed in 95 patients was investigated for the prognostic value of clinical, histological and DNA cytophotometric parameters. Metastases almost exclusively occurred among ileal carcinoids, which also were responsible for all 14 cases of lethal outcome observed during the follow-up period of mean 42 months. Aneuploid DNA values could be determined significantly more frequently among ileal than in non-ileal carcinoids and showed - upon analysis of the total group of gastrointestinal neuroendocrine tumours - a significant correlation to lethal course of disease. In addition, among 18 cases with primary and secondary carcinoid manifestations available for DNA cytophotometry, an association between the DNA content of metastatic neuroendocrine tumours and prognosis came to light. When applied to the group of ileal neoplasms, however, the parameter DNA content did not allow a better prognostic assessment.

[Diagnostic guidelines for adrenal gland nodules: typing, assessment of biological potential and prognosis, molecular pathogenesis]. / Diagnostische Leitlinien bei Knoten in der Nebenniere: Typisierung, Dignitäts- und Prognoseerfassung, molekulare Pathogenese.

The aim of morphological tumour diagnosis is to answer clinical questions on type, biological potential, prognosis and aetiology of individual neoplasms. The limitations and perspectives of different methods used in the diagnosis of adrenal tumours, ranging from histology to molecular genetic DNA analyses, are described. When surgical specimens from adrenal neoplasms cannot be typed on the basis of histology and/or with clinica data (e.g. endocrine symptoms and history) as adrenocortical tumours, phaeochromocytomas or metastases to the adrenal, immunohistological investigations with a panel of different antibodies are necessary. After identification of the tissue derivation of an individual adrenal tumour, its biological potential must be assessed. Among adrenocortical neoplasms, adenomas and carcinomas can be distinguished by evaluation of various histological parameters (including structural features and signs of invasion) according to defined algorithms. In addition, conventional histology (by estimation of mitotic activity) allows the discrimination of tumours with especially high malignant potential from other adrenocortical carcinomas. In contrast, among adrenomedullary tumours even the combined use of histological, immunohistological and DNA cytophotometric techniques only allows the definition of risk groups (benign versus suggestive of malignancy), while reliable recognition of an individual malignant phaeochromocytoma is so far impossible. The question as to whether a particular phaeochromocytoma represents a sporadic tumour or a neoplasm inherited as one feature of a defined syndrome cannot be answered with the above methods, but only by the application of molecular genetic techniques.

[Cytokeratin expression of benign and malignant epithelial thyroid gland tumors. An immunohistologic study of 154 neoplasms using 8 different monoclonal cytokeratin antibodies]. / Zytokeratinexpression benigner und maligner epithelialer Schilddrüsentumoren. Eine immunohistologische Untersuchung an 154 Neoplasien unter Einsatz von 8 verschiedenen monoklonalen Zytokeratinantikörpern.

Using 8 different monoclonal antibodies, immunohistology was performed on 36 follicular adenomas and on 28 follicular, 34 papillary, 27 medullary and 29 anaplastic carcinomas of the thyroid. The panel of antibodies was directed against broad-spectrum cytokeratins (pan-CK, antibody lu-5), against basic and acid high-molecular-weight CK of types #1, 5, 10 and 14, against basic (#5 and 6) and acid high-molecular-weight CK (#13) and against basic (#7 and #8) and acid low-molecular-weight CK (#19 and #20). With the exception of a large number of anaplastic carcinomas, nearly all other tumours exhibited strong immunoreactivity with antibodies against pan-CK, CK 8 and CK 19. CK 20 expression was exclusively shown for 2 medullary carcinomas. Reactivity for high-molecular-weight CK could only, each time focally, be demonstrated for 14 papillary and 2 follicular carcinomas and for 2 anaplastic carcinomas with partial squamous differentiation. Thirteen anaplastic carcinomas were not decorated by any of the CK antibodies applied. CK 7 staining exceeding the staining of individual cells was observed in 26 papillary cancers. In contrast, such a finding could only be obtained with each one follicular adenoma, medullary carcinoma and anaplastic carcinoma and with 5 follicular carcinomas. These results confirm earlier studies in that CK 20 expression among thyroid tumours is restricted to the neuroendocrine medullary carcinomas and that in a larger percentage of anaplastic thyroid carcinomas an epithelial phenotype can not be demonstrated even upon using broad-spectrum CK antibodies. New is the finding that there exist considerable differences between papillary carcinomas and all other non-papillary thyroid tumours regarding CK 7 expression. This result might be of differential diagnostic value for the distinction of follicular and papillary thyroid neoplasias which sometimes have an overlapping histological pattern.

DNA cytophotometry and prognosis in typical and atypical bronchopulmonary carcinoids. A clinicomorphologic study of 100 neuroendocrine lung tumors.

Surgical material obtained from 100 patients with typical carcinoids (TC) and atypical carcinoids (AC) of the lung (including 100 primary, four residual tumors, and four lymph node or distant metastases) was investigated by conventional histology and scanning DNA cytophotometry. Of the 60 TC (96%), 58 exhibited euploid DNA histograms compared with only 20 (50%) of the 40 AC. The morphologic findings were related to the patients' survival (median observation period, 9 years). Statistical analyses disclosed the histologic type of disease (TC versus AC) and the DNA content of tumors (euploid versus aneuploid) to affect prognosis significantly (p < 0.001). Deaths resulting from tumor were exclusively observed among patients with atypical (eight of 40) or DNA aneuploid carcinoids (eight of 22). Six patients were alive with persistent tumor manifestations 3 to 20 years after initial diagnosis, four with DNA diploid primary carcinoids. The presence of lymph node metastases alone was not associated with poor prognosis as long as the primary tumor or the related metastases showed a diploid DNA content. DNA cytophotometry thus might be regarded as an adjunctive prognostic criterion in individual carcinoid cases.

Prognostic significance of nuclear DNA content and proliferative activity in renal cell carcinomas. A clinicopathologic study of 58 patients using mitotic count, MIB-1 staining, and DNA cytophotometry.

BACKGROUND: For a variety of human malignancies, static DNA cytophotometry and immunostaining for the Ki-67 antigen using the antibody MIB-1 have provided significant prognostic information. METHODS: Surgical specimens of 58 renal cell carcinomas (RCCs) were investigated by conventional histology, DNA cytophotometry, and MIB-1 immunostaining. RESULTS: The MIB-1 indices and DNA data were found not only to be significantly correlated with various other morphologic parameters, but also to the clinical behavior of RCC. In the course of this study (median observation period: 31 months), 27% of patients died from RCC. None of these patients belonged to the group of 37 patients with RCCs exhibiting diploid or euploid DNA histograms. Lethal outcome occurred in only 16 of the 21 patients (76%) with noneuploid or aneuploid histogram tumors (P < 0.0001). According to their MIB-1 indices and upon choosing different cutoff levels, the 58 RCCs were categorized into 2 groups with either low or high proliferative activity. Using the median and the mean MIB-1 index as cutoffs, none of the patients with tumors showing low proliferative activity had died, whereas 16 of 29 patients (55%) or, respectively, 16 of 25 patients (64%) with tumors exhibiting high proliferative activity, had died from RCC (P < 0.0001). CONCLUSIONS: In addition to tumor grade and stage, both a high MIB-1 index and a noneuploid or aneuploid DNA histogram of a given RCC have the potential to identify tumor patients with an impaired prognosis.

Central neurocytoma: clinical, immunohistologic, and biologic findings of a human neuroglial progenitor tumor.

BACKGROUND: Central neurocytomas are rare brain tumors recognized by their typical radiologic and histologic features. In general, a good prognosis is achieved by total removal. The histogenesis is still under debate, but a neuronal origin is widely assumed. METHODS: This study presents the clinical and immunohistologic findings of five patients and the results of cell culture experiments of two patients with central neurocytoma treated surgically between 1983 and 1993. RESULTS: The patient age at diagnosis ranged from 21 to 30 years (mean, 25 years). The male-to-female ration was 1:4. Raised intracranial pressure due to hydrocephalus was the main cause of the clinical manifestations. Total resection was achieved in two cases. Four patients received radiotherapy. One patient suffered a recurrence 1 year after surgery, requiring a second resection and radiotherapy. Follow-up studies took place between 1 and 10.5 years (mean, 7.1 years). To date, all patients are free of their tumors. Two patients suffered from permanent memory disturbances after surgery. Immunohistochemistry confirmed the neuronal nature of the tumors. Cell-culture studies, which have been carried out for the first time, demonstrated concomitant expression of neuronal (synaptophysin) and glial (GFAP) markers. CONCLUSION: Total removal is the therapy of choice. In tumor recurrence or limited surgery (e.g. due to severe affliction of the fornical structures), radiotherapy has shown to be efficacious. The cell-culture experiments give new insight on the histogenesis of central neurocytoma, indicating that the tumor arises from an undifferentiated precursor cell with the capacity of bipotential neuroglial differentiation.

Absence of RET proto-oncogene point mutations in sporadic hyperplastic and neoplastic lesions of the parathyroid gland.

We investigated the possible role of RET proto-oncogene mutations in the development of sporadic hyperplastic, benign, and malignant parathyroid lesions. DNA extracted from paraffin-embedded specimens of forty parathyroid lesions was screened for RET proto-oncogene point mutations in exons 10, 11, and 16 by nonisotopic polymerase chain reaction-based single-strand conformation polymorphism and heteroduplex gel electrophoresis. The nucleotide sequence of samples with aberrant band patterns was identified by nonisotopic direct sequencing of polymerase chain reaction-amplified DNA. Parathyroids of seven patients with multiple endocrine neoplasia type 2A (MEN 2A) and MEN 2B served as positive controls. None of the eight hyperplastic lesions, three cases of parathyromatosis, ten parathyroid adenomas, eleven carcinomas or one normal parathyroid gland contained mutations in each of the three RET exons tested. Six MEN-2A-associated hyperplastic glands exhibited identical band shifts in the polymerase chain reaction single-strand conformation polymorphism analysis of exon 11, which corresponded to a Cys 634-->Arg substitution in the nucleotide sequence analysis (TGC-->CGC), whereas in the MEN 2B parathyroid specimen a point mutation was found at codon 918 of exon 16 (ATG-->ACG), causing a Met 918-->Thr substitution. Our data indicate that RET mutations of the MEN 2 loci in exons 10, 11, and 16 are not involved in the development of sporadically occurring benign or malignant parathyroid lesions. Furthermore, our results are in accordance with the observation that MEN 2A patients with Cys 634-->Arg (germline) mutations have a higher risk of developing parathyroid disease than those with other mutations at codon 634.

[Morphological typing, evaluation of tumor dignity and prognosis and etiologic classification of adrenomedullary and adrenocortical neoplasias]. / Morphologische Typisierung, Dignitts- und Prognose-beurteilung sowie ätiologische Einordnung adrenomedullärer und adrenokortikaler Neoplasien.

The aim of morphological tumour diagnosis is to answer clinical questions on type, biological potential, prognosis and aetiology of individual neoplasms. The limitations and perspectives of different methods used in the diagnosis of adrenal tumours, ranging from histology to molecular genetic DNA analyses, are described. When surgical specimens from adrenal neoplasms cannot be typed on the basis of histology and/or with clinica data (e.g., endocrine symptoms and history) as adrenocortical tumours, phaeochromocytomas or metastases to the adrenal, immunohistological investigations with a panel of different antibodies are necessary. After identification of the tissue derivation of an individual adrenal tumour, its biological potential must be assessed. Among adrenocortical neoplasms, adenomas and carcinomas can be distinguished by evaluation of various histological parameters (including structural features and signs of invasion) according to defined algorithms. In addition, conventional histology (by estimation of mitotic activity) allows the discrimination of tumours with especially high malignant potential from other adrenocortical carcinomas. In contrast, among adrenomedullary tumours even the combined use of histological, immunohistological and DNA cytophotometric techniques only allows the definition of risk groups (benign versus suggestive of malignancy), while reliable recognition of an individual malignant phaeochromocytoma is so far impossible. The question as to whether a particular phaeochromocytoma represents a sporadic tumour or a neoplasm inherited as one feature of a defined syndrome cannot be answered with the above methods, but only by the application of molecular genetic techniques.

Secretoneurin in bronchopulmonary carcinoids--immunohistochemical comparison with chromogranins A and B and secretogranin II.

Ninety-nine classical and 11 atypical bronchopulmonary carcinoids were investigated immunohistochemically with an antibody against secretoneurin, a peptide proteolytically processed from secretogranin II (chromogranin C), as well as antibodies against chromogranin A and B and secretogranin II. Secretoneurin was immunolocalized in 86 tumours (78 classical and eight atypical carcinoids); secretogranin II was found in the same tumours in a similar distribution, whereas chromogranin A was present in all 100 and chromogranin B in 106 tumours investigated. Bronchopulmonary carcinoids are usually not associated with clinically or biochemically distinct syndromes. Although we found bronchial carcinoids with different immunohistochemical chromogranins/secretogranin patterns, no correlation with the biological behaviour of these tumours could be demonstrated.

Multiple endocrine neoplasia type 1 (MEN 1) revisited.

Multiple endocrine neoplasia type 1 (MEN 1) is an inherited disease of the neuroendocrine cell system affecting primarily the parathyroids, pancreas, duodenum and the anterior pituitary. The pancreatic and duodenal tumours may metastasize, but generally have a low malignant potential. The diagnosis of MEN 1 is usually made in the second decade of life and based on the involvement of at least two organs and a family history. The recent discovery of the MEN 1 locus on the centromeric region of the long arm of chromosome 11 may become a further diagnostic criterion. The use of flanking DNA markers permits presymptomatic testing for MEN 1 in affected families.

[Lymphoepithelial carcinoma of the thyroid gland. A thyroid gland carcinoma with thymus-like differentiation]. / Lymphoepitheliales Karzinom der Schilddrüse. Ein Schilddrüsenkarzinom mit thymusartiger Differenzierung.

An example of lymphoepithelial carcinoma of the thyroid is reported, which was first described 10 years ago and has not as yet been documented in the German literature. This tumour is morphologically identical to the lymphoepithelioma of the nasopharynx. In contrast to neoplasms of such morphology occurring in a variety of other organs, the acronym CASTLE--for carcinoma showing thymus-like differentiation--has been proposed by Chan and Rosai for lymphoepithelioma-like carcinomas of the thyroid gland. In this paper, the aforenamed authors' concept of the histogenesis of thyroid tumours with thymic differentiation is presented.