S. Synoracki

NGS based identification of mutational hotspots for targeted therapy in anaplastic thyroid carcinoma

Context Anaplastic thyroid carcinoma (ATC) represents one of the most aggressive carcinomas with no consistent survival benefit when treated with conventional radiochemotherapy. Approaches targeting “oncogene addiction” of ATC are increasingly explored and first promising results have been reported in single case studies. Objective To determine the prevalence of mutations in known thyroid oncogenes and signalling pathways amendable to targeted therapy in a large cohort of ATC. Results In 118 ATC (57 male/ 61 female) a total of 165 mutations were found. Genes involved in the MAPK/ERK and PI3K pathway (BRAF 11.0%, HRAS 4.2%, KRAS 7.6%, NRAS 7.6%, PI3KCA 11.8%) were altered in 33%. Targetable receptor tyrosine kinases were mutated in 11%. The most frequently altered genes were TERT in 86/118 (73%) and p53 in 65/118 (55%) cases. No mutations were found analysing ALK, KIT, MET and mTOR. Materials and Methods Next generation sequencing (NGS) was performed in FFPE samples from 118 ATC using MiSeq (Illumina) and CLC Cancer Research Workbench (CLCbio; Qiagen) for mutation analysis in: ALK, BRAF, CDKN2A, EGFR, ERBB2, HRAS, KIT, KRAS, MET, mTOR, NRAS, PDGFRA, PI3KCA, p53, RB1, RET and TSC2. Sanger sequencing was used to detect TERT promotor mutations. Conclusions To our knowledge this is the largest study analysing mutations for targeted therapy of ATC. We found that 33% of ATC harbour mutations in pathways amendable to targeted therapy. Molecular screening in ATC is suggested for targeted therapies since current conventional treatment for ATC proved mainly futile.

[Thyroid C cells and their pathology: Part 1: normal C cells, - C cell hyperplasia, - precursor of familial medullary thyroid carcinoma].

The C cells (parafollicular) of the human thyroid gland are predominantly located within the thyroid follicles, are of neuroendocrine origin and produce and secrete the peptide hormone calcitonin. Calcitonin is clinically utilized as a screening marker to detect occult medullary thyroid carcinoma (MTC) as well as in the follow-up of patients with MTC. An increase in the number of C cells is designated as C cell hyperplasia (CCH). Neoplastic CCH is caused by an autosomal dominant inherited mutation of the RET protooncogene, which develops into invasive familial MTC in the setting of multiple endocrine neoplasia (MEN) type 2 depending on the location of the mutation in the RET gene with a high variation in latency. According to the current state of knowledge CCH without a germline mutation in the RET protooncogene, designated as non-MEN2-associated CCH, seems to be unrelated to the development of sporadic MTC.

Die C-Zellen der Schilddrüse und ihre Pathologie

The C cells (parafollicular) of the human thyroid gland are predominantly located within the thyroid follicles, are of neuroendocrine origin and produce and secrete the peptide hormone calcitonin. Calcitonin is clinically utilized as a screening marker to detect occult medullary thyroid carcinoma (MTC) as well as in the follow-up of patients with MTC. An increase in the number of C cells is designated as C cell hyperplasia (CCH). Neoplastic CCH is caused by an autosomal dominant inherited mutation of the RET protooncogene, which develops into invasive familial MTC in the setting of multiple endocrine neoplasia (MEN) type 2 depending on the location of the mutation in the RET gene with a high variation in latency. According to the current state of knowledge CCH without a germline mutation in the RET protooncogene, designated as non-MEN2-associated CCH, seems to be unrelated to the development of sporadic MTC.

Comparison of the PI3KCA pathway in circulating tumor cells and corresponding tumor tissue of patients with metastatic breast cancer

The aim of the present study was to compare the phosphatidylinositol 3-kinase (PI3KCA)-AKT serine/threonine kinase (AKT) pathway in circulating tumor cells (CTCs) and corresponding cancerous tissues. Stemness-like circulating tumor cells (slCTCs) and CTCs in epithelial-mesenchymal transition (EMT) have been implicated as the active source of metastatic spread in breast cancer (BC). In this regard, the PI3KCA-AKT signaling pathway was demonstrated to be implicated in and to be frequently mutated in BC. The present study compared this pathway in slCTCs/CTCs in EMT and the corresponding tumor tissues of 90 metastatic BC patients (pts). slCTCs and CTCs in EMT were isolated using the AdnaTest EMT-1/StemCell for the detection of aldehyde dehydrogenase 1 family member A1 (ALDH1) (singleplex PCR) and PI3KCA, AKT2 and twist family bHLH transcription factor 1 (multiplex PCR). Tumor tissue was investigated for PI3KCA hotspot mutations using Sanger sequencing of genomic DNA from micro-dissected formalin-fixed paraffin-embedded tissue, and for the expression of ALDH1 and phosphorylated AKT (pAKT), and phosphatase and tensin homolog (PTEN) loss, by immunohistochemistry. slCTCs were identified in 23% of pts (21/90 pts) and CTCs in EMT in 56% (50/90 pts) of pts. pAKT and ALDH1 positivity in tumor tissue was identified in 47 and 9% of cases, respectively, and a PTEN loss was observed in 18% of pts. A significant association was detected between pAKT expression in cancerous tissue and AKT2 expression in CTCs (P=0.037). PI3KCA mutations were detected in 32% of pts, most frequently on exons 21 (55%) and 10 (45%). Pts with PI3KCA mutations in tumor tissue had a significantly longer overall survival than pts with wild-type PI3KCA expression (P=0.007). Similar results were obtained for pts with aberrant PI3KCA signaling in CTCs and/or aberrant signaling in cancerous tissue (P=0.009). Therapy-resistant CTCs, potentially derived from the primary tumor or metastatic tissue, may be eliminated with specific PI3K pathway inhibitors, alone or in combination, to improve the prognosis of metastatic BC pts.

Intraoperativer Gefrierschnitt der Schilddrüse

The goal of evaluation of intraoperative frozen sections of the thyroid gland is to achieve a definitive diagnosis which determines the subsequent surgical management as fast as possible; however, due to the specific methodological situation of thyroid frozen sections evaluation a conclusive diagnosis can be made in only some of the cases. If no conclusive histological diagnosis is possible during the operation, subsequent privileged processing of the specimen allows a final diagnosis at the latest within 48 h in almost all remaining cases. Applying this strategy, both pathologists and surgeons require a high level of communication and knowledge regarding the specific diagnostic and therapeutic peculiarities of thyroid malignancies because different surgical strategies must be employed depending on the histological tumor subtype.

Entzündungen der Schilddrüse

This review article deals with the classification, clinical features and morphology of thyroiditis. These inflammatory diseases account for approximately 20 % of all thyroid diseases. The vast majority of cases of thyroiditis are of immunogenic origin while non-immunogenic thyroiditis (caused by pathogens or iatrogenic) is a rarity.

Entzündungen der Nebenschilddrüsen

Inflammation of the parathyroid glands is rare when compared to other endocrine organs. This leads to the use of descriptive terms as well as the lack of a generally accepted classification for inflammatory disorders of the parathyroid glands. This review article proposes that parathyroid inflammation be subdivided morphologically into (a) non-specific lymphocytic infiltration, which is more an expression of damage to small vessels, due to e. g. severe systemic inflammation or myocardial infarction, (b) autoimmunogenic lymphocytic parathyroiditis, (c) nonimmunogenic inflammation caused by granulomatous diseases or infections and (d) invasive sclerosing (peri) parathyroiditis. As only parathyroid glands removed due to hyperparathyroidism and normal parathyroid glands incidentally removed during thyroid surgery are seen almost exclusively in routine histopathology, virtually no information about the morphological correlate of hypoparathyroidism is available.ZusammenfassungEntzündungen der Nebenschilddrüsen sind im Vergleich mit anderen endokrinen Organen selten. Dies führt sowohl zur Verwendung deskriptiver Begriffe als auch zum Fehlen einer allgemein anerkannten Klassifikation. Im vorliegenden Übersichtsartikel wird vorgeschlagen, die Entzündungen der Nebenschilddrüse morphologisch in (a) eine unspezifische lymphozytäre Infiltration, die eher Ausdruck einer allgemeinen Schädigung kleiner Gefäße bei z. B. schweren Entzündungen oder Herzinfarkt ist, (b) eine autoimmunogen bedingte lymphozytäre Parathyreoiditis, (c) nichtimmunogene Entzündungen bei granulomatösen Erkrankungen und Infektionen und (d) die invasiv-sklerosierende (Peri-)Parathyreoiditis einzuteilen. Da im histologischen Untersuchungsgut praktisch ausschließlich Nebenschilddrüsen bei Hyperparathyreoidismus und normale Nebenschilddrüsen, die unabsichtlich bei Schilddrüsenoperationen mitentfernt wurden, gesehen werden, gibt es so gut wie keine Informationen zum morphologischen Korrelat des Hypoparathyreoidismus.AbstractInflammation of the parathyroid glands is rare when compared to other endocrine organs. This leads to the use of descriptive terms as well as the lack of a generally accepted classification for inflammatory disorders of the parathyroid glands. This review article proposes that parathyroid inflammation be subdivided morphologically into (a) non-specific lymphocytic infiltration, which is more an expression of damage to small vessels, due to e. g. severe systemic inflammation or myocardial infarction, (b) autoimmunogenic lymphocytic parathyroiditis, (c) nonimmunogenic inflammation caused by granulomatous diseases or infections and (d) invasive sclerosing (peri) parathyroiditis. As only parathyroid glands removed due to hyperparathyroidism and normal parathyroid glands incidentally removed during thyroid surgery are seen almost exclusively in routine histopathology, virtually no information about the morphological correlate of hypoparathyroidism is available.

Klinische Bedeutung der Schilddrüsenzytologie

The cytological evaluation of fine needle biopsies (FNB) of the thyroid gland crucially depends on a close cooperation between clinicians and cytopathologists. Scintigraphy, sonography as well as clinical data and patient history are necessary for a correct interpretation of the indications for FNB; moreover, these data are of outstanding importance for cytopathologists for the correct interpretation of the cytomorphological findings. This overview describes the present standards in the acquisition, technical workup and cytopathological interpretation of thyroid gland tissue obtained by FNB, particularly focusing on the rapidly growing relevance of additional molecular pathological investigations to increase the diagnostic accuracy of thyroid FNB.ZusammenfassungDie zytologische Beurteilung der Feinnadelbiopsie (FNB) der Schilddrüse setzt eine enge Kommunikation von Klinikern und Zytopathologen voraus. Szintigraphie, Sonographie und klinisch-anamnestische Befunde sind zur klinischen Indikationsstellung für die Punktion unerlässlich; die Kenntnis dieser Befunde ist aber auch für den Pathologen zur abschließenden Interpretation des zytomorphologischen Befundes unerlässlich. Der vorliegende Übersichtsartikel beschreibt den gegenwärtigen Standard der Gewinnung, Verarbeitung und zytopathologischen Beurteilung von Schilddrüsenpunktaten. Molekularpathologischen Zusatzuntersuchungen am zytologischen Material kommt eine zunehmende Bedeutung bei der präzisen Befundkategorisierung zu.AbstractThe cytological evaluation of fine needle biopsies (FNB) of the thyroid gland crucially depends on a close cooperation between clinicians and cytopathologists. Scintigraphy, sonography as well as clinical data and patient history are necessary for a correct interpretation of the indications for FNB; moreover, these data are of outstanding importance for cytopathologists for the correct interpretation of the cytomorphological findings. This overview describes the present standards in the acquisition, technical workup and cytopathological interpretation of thyroid gland tissue obtained by FNB, particularly focusing on the rapidly growing relevance of additional molecular pathological investigations to increase the diagnostic accuracy of thyroid FNB.

Inflammatory diseases of the thyroid gland

This review article deals with the classification, clinical features and morphology of thyroiditis. These inflammatory diseases account for approximately 20 % of all thyroid diseases. The vast majority of cases of thyroiditis are of immunogenic origin while non-immunogenic thyroiditis (caused by pathogens or iatrogenic) is a rarity.

Entzündungen der Schilddrüse@@@Inflammatory diseases of the thyroid gland

This review article deals with the classification, clinical features and morphology of thyroiditis. These inflammatory diseases account for approximately 20 % of all thyroid diseases. The vast majority of cases of thyroiditis are of immunogenic origin while non-immunogenic thyroiditis (caused by pathogens or iatrogenic) is a rarity.