Toll-like receptor 5 and 7 expression may impact prognosis of HPV-positive oropharyngeal squamous cell carcinoma patients

A large subset of oropharyngeal squamous cell carcinomas (OPSCCs) is associated with HPV infection and has better outcome than non-viral-related tumors. Various malignancies also carry a role for TLRs, key activators of inflammation and innate immunity. We examined the expression of TLRs in OPSCC, and their association with HPV status and treatment outcome. TLR 5, 7, 9, and p16 were studied by immunohistochemistry and HPV status was detected with in situ hybridization in 202 tumors of consecutively treated OPSCC patients using tissue microarray method. The relations between TLR expression and HPV status, p16 expression, clinicopathological factors, and survival were analyzed. TLR 5, 7, and 9 expression patterns differed between HPV-positive and -negative tumors, and they were statistically significantly associated with history of smoking, heavy drinking, tumor site, grade, size (T), metastasis (N), and stage. Moreover, in HPV-positive tumors the expression of TLR 5 and 7 correlated with tumor recurrence. After adjustment, among HPV-positive OPSCC patients, high TLR 5 and low TLR 7 expression were associated with poor disease-specific survival. Our results indicate that TLR 5 and 7 may have a role in the prognostication of HPV-positive OPSCC, however, further studies are needed to clarify the comprehensive role of these TLRs in OPSCC.Peer reviewe

PCSK2 expression in neuroendocrine tumors points to a midgut, pulmonary, or pheochromocytoma-paraganglioma origin

Neuroendocrine tumors (NETs) are often diagnosed from the metastases of an unknown primary tumor. Specific immunohistochemical (IHC) markers indicating the location of a primary tumor are needed. The proprotein convertase subtilisin/kexin type 2 (PCSK2) is found in normal neural and neuroendocrine cells, and known to express in NETs. We investigated the tissue microarray (TMA) of 86 primary tumors from 13 different organs and 9 metastatic NETs, including primary tumor-metastasis pairs, for PCSK2 expression with polymer-based IHC. PCSK2 was strongly positive in all small intestine and appendiceal NETs, the so-called midgut NETs, in most pheochromocytomas and paragangliomas, and in some of the typical and atypical pulmonary carcinoid tumors. NETs showing strong positivity were re-evaluated in larger tumor cohorts confirming the primary observation. In the metastases, the expression of PCSK2 mirrored that of the corresponding primary tumors. We found negative or weak staining in NETs from the thymus, gastric mucosa, pancreas, rectum, thyroid, and parathyroid. PCSK2 expression did not correlate with Ki-67 in well-differentiated NETs. Our data suggest that PCSK2 positivity can indicate the location of the primary tumor. Thus, PCSK2 could function in the IHC panel determined from screening metastatic NET biopsies of unknown primary origins.Peer reviewe

Treponema denticola chymotrypsin-like protease as associated with HPV-negative oropharyngeal squamous cell carcinoma

BACKGROUND: An opportunistic oral pathogen, Treponema denticola (Td), has been linked to orodigestive carcinogenesis, but its role in oropharyngeal squamous cell carcinoma (OPSCC) has remained open. We evaluated the presence of Td chymotrypsin-like protease (Td-CTLP) in a series of 201 unselected consecutive OPSCC patients, and the relation of the Td-CTLP to human papillomavirus (HPV) status, to expression of toll-like receptors (TLR) 5, 7, and 9, and to clinical parameters and patient outcome. METHODS: Clinicopathological data came from hospital registries. The expression of cell surface-bound Td-CTLP was evaluated by immunohistochemistry. Immunoexpression of TLRs 5, 7, and 9, and HPV status we studied earlier in this patient series. RESULTS: We detected Td-CTLP in 81% of the OPSCC, and especially in HPV-negative tumours (48% of all OPSCCs). Among the HPV-positive tumours (52% of all OPSCCs), low Td-CTLP expression associated with low TLR 5 and high TLR 7 expression. Among those HPV-negative, higher TLR 5 and lower TLR 7 expression associated with high Td-CTLP expression. Strong Td-CTLP expression associated with poor disease-specific survival, but no similar association among HPV-positive and HPV-negative subgroups emerged. CONCLUSIONS: Td-CTLP was highly expressed in OPSCC and was associated with the HPV status of tumour tissue.Peer reviewe

Neuroendocrine tumors : the quality of prognostic and predictive markers

Neuroendocrine neoplasms (NENs), rare tumors derived from disseminated neuroendocrine (NE) cells, can develop in different organs. NENs are sometimes diagnosed from metastatic tissue. The primary tumor location and grade affect both prognosis and the management of the disease. Histomorphologically, NENs show similarities independent of tumor origin. Immunohistochemistry (IHC) plays a key role in NEN diagnostics. Chromogranin A and synaptophysin expression confirms the NE nature of tumor cells, while tissue-specific IHC markers may indicate the primary tumor location. The proliferation marker Ki-67 determines the tumor grade of an NEN. Careful verification and validation of diagnostic, prognostic, and predictive markers, as well as reliable and reproducible staining assessment are necessary in NEN diagnostics. This thesis primarily aimed to identify new IHC markers for NENs and evaluate scoring methods for Ki-67 stainings using digital pathology and image analysis. The studies relied on a NE tumor (NET) cohort consisting of 91 NENs from 12 different primary sites, including primary–metastasis pairs. Tissue microarray (TMA) blocks were constructed from archived tumor specimens. First, the specificity of the primary antibodies studied was confirmed through technical validation, after which each marker’s expression was evaluated. The Ki-67 proliferation index determining the NET grade was examined using the image analysis software ImmunoRatio and conventional eyeballing. All NENs expressed neuropeptide S receptor 1 (NPSR1) and the neuropeptide S (NPS) ligand except for pheochromocytomas (PHEOs). The expression of proprotein convertase subtilisin/kexin type 2 (PCSK2) was identified in midgut NENs, PHEOs, paragangliomas, and pulmonary carcinoids. NENs from other primary locations were PCSK2-negative. The staining profiles of NPSR1, NPS, and PCSK2 were similar in primary tumors compared with metastatic samples. Validation of somatostatin receptor (SSTR) primary antibody clones revealed that only a few localized the receptor to the correct cell components. UMB clones exhibited the best cell membrane positivity. Different NENs showed varying SSTR 1-5 profiles depending on the primary location and tumor grade. In addition, assessment of the Ki-67 index using eyeballing revealed more variation than computer-assisted scoring. Thus, grading using image analysis proved more reliable than traditional analysis. To conclude, NPSR1 and PCSK2 represent potential markers to identify NETs and the possible primary tumor origin. Standardized IHC methods and the choice of primary antibody clone remain crucial for reliably analyzing diagnostic markers, while computer-aided proliferation index assessment yields a more objective classification of NENs.Neuroendokriiniset (NE) kasvaimet muodostavat heterogeenisen kasvainryhmän, joilla on yhtenäisiä histomorfologisia piirteitä valomikroskopiassa. Nämä diffuusista neuroendokriinisolujen järjestelmästä lähtöisin olevat kasvaimet ovat harvinaisia ja niitä esiintyy eri puolilla elimistöä. Neuroendokriinisten kasvainten ilmaantuvuus on nousussa. Kliinisesti vähäoireiset NE-kasvaimet diagnosoidaan välillä metastaasien perusteella. Kasvaintyypistä, primaarikasvaimen sijainnista ja erilaistumisasteesta riippuen taudin ennuste sekä hoitokäytännöt vaihtelevat. Immunohistokemiallisten värjäysten avulla tarkennetaan neuroendokriinisen kasvaimen histologista luonnetta, pyritään selvittämään levinneen kasvaimen emotuumoria ja tarkastellaan ennusteeseen sekä hoitoihin vaikuttavia tekijöitä. Immunohistokemialla onkin vahva rooli diagnostiikassa. Tämän väitöskirjatyön tavoite oli löytää uusia neuroendokriinisia kudosmerkkiaineita ja huomioida samalla immunohistokemiallisten menetelmien laadulliset näkökulmat. Lisäksi tarkasteltiin Ki-67 värjäyksen tulkintatapaa ja sen vaikutusta kasvainten Ki-67 pohjaiseen luokitteluun. Kasvainaineisto koostui neuroendokriinisista kasvaimista, jotka olivat lähtöisin 12 eri elimestä, sisältäen myös primaari-metastaasi pareja. Kasvainmateriaalista valmistettiin monikudosblokki (tissue microarray). Tutkittavien merkkiaineiden ilmentymistä ja Ki-67 proliferaatioindeksin tulkintaa analysointiin tämän materiaalin avulla. Tutkituista immunohistokemiallisista merkkiaineista neuropeptidi S reseptori (NPSR1) ja ligandi neuropeptide S (NPS) ilmentyivät kaikissa neuroendokriinikasvaintyypeissä, paitsi feokromosytoomissa. Entsyymimarkkeri proprotein convertase subtilisin/kexin type 2 (PCSK2) ilmentyi vain tietyissä neuroendokriinikasvaimissa: keskisuolen kasvaimissa, feokromosytoomissa, paraganglioomissa ja keuhkon karsinoideissa. Somatostatiinireseptori vasta-aineiden immunohistokemiallisessa validaatiossa havaittiin, että värjäytyminen on klooni sidonnaista, ja vain tietyillä klooneilla saavutettiin kohde antigeenille ominainen membraanivärjäytyminen. Ki-67-proliferaatioindeksin eri määrittämistavoilla oli vaikutusta NE-kasvaimen gradukseen. Kuva-analyysi pystyi merkitsevästi tasalaatuisempaan Ki-67 analyysiin verrattuna perinteiseen subjektiiviseen tulkitaan. Merkkiaineiden NPSR1 ja NPS positiivisuus tukee kasvaimen NE alkuperää. PCSK2 positiivisuus puolestaan antaa viitteitä mahdollisesta emotuumorin sijainnista. Immunohistokemiallisten merkkiaineiden validaatio on tärkeää varmistaen tarkasteltavien antigeenien oikean sijainnin ja tulkinnan. Tietokoneavusteinen Ki-67 analysointi parantaa NE-kasvainten luokittelun objektiivisuutta

Evidence of a causal effect of genetic tendency to gain muscle mass on uterine leiomyomata

Uterine leiomyomata (UL) are the most common tumours of the female genital tract and the primary cause of surgical removal of the uterus. Genetic factors contribute to UL susceptibility. To add understanding to the heritable genetic risk factors, we conduct a genome-wide association study (GWAS) of UL in up to 426,558 European women from FinnGen and a previous UL meta-GWAS. In addition to the 50 known UL loci, we identify 22 loci that have not been associated with UL in prior studies. UL-associated loci harbour genes enriched for development, growth, and cellular senescence. Of particular interest are the smooth muscle cell differentiation and proliferation-regulating genes functioning on the myocardin-cyclin dependent kinase inhibitor 1A pathway. Our results further suggest that genetic predisposition to increased fat-free mass may be causally related to higher UL risk, underscoring the involvement of altered muscle tissue biology in UL pathophysiology. Overall, our findings add to the understanding of the genetic pathways underlying UL, which may aid in developing novel therapeutics.Peer reviewe