Az urogenitális daganatok molekuláris klasszifikációja

The molecular genetic dogma is valid that the histological variants of a given cancer represent genetic variants. Basis of this subclassification is known in clear cell renal cancer but still a mistery in prostate or bladder cancers. Meanwhile another genetic dogma developed recently that a given histological variant of a cancer can further be subdivided based on molecular characteristics. Best examples are clear cell renal cancer, adenocarcinoma of the prostate or transitional cell carcinoma of the bladder. This new knowledge helps in the differential diagnostics of cancer, and in determining prognosis, but also provides an opportunity to better tailor existing therapies even to consider novel target agents. Discovery of the molecular subtypes of cancers (such as leukemia or lung adenocarcinoma) contributed significantly to the extension of the progression-free or overall survival of cancer patients, and it is expected that it could lead to similar effects in case of urogenital cancers

A patológiai diagnosztika jelentősége a daganatos betegségek személyre szabott orvoslásában

Due to the developments of pathology in the past decades (immunohistochemistry and molecular pathology) classification of cancers changed fundamentally, laying a ground for personalized management of cancer patients. Our picture of cancer is more complex today, identifying the genetic basis of the morphological variants. On the other hand, this picture has a much higher resolution enabling us to subclassify similar histological cancer types based on molecular markers. This redefined classification of cancers helps us to better predict the possible biological behavior of the disease and/or the therapeutic sensitivity, opening the way toward a more personalized treatment of this disease. The redefined molecular classification of cancer may affect the universal application of treatment protocols. To achieve this goal molecular diagnostics must be an integral and reimbursed part of the routine pathological diagnostics. On the other hand, it is time to extend the multidisciplinary team with molecular pathologist to improve the decision making process of the management of cancer patients

A tüdőrák molekuláris patológiai vizsgálatának feltételei

From the aspect of the contemporary pathologic diagnostics of lung cancer the tissue obtained is a key issue since small biopsies and cytology still play a major role. In the non-small cell lung cancer era cytology is considered equal to biopsy however, in recent years it is unable to provide quality diagnosis and must be replaced by biopsy. Various molecular techniques can handle various different tissue samples which must be considered during molecular pathology diagnosis. Moreover, tumor cell-normal cell ratio in the obtained tissue, as well as the absolute tumor cell number have great significance, which information must be provided in the primary lung cancer diagnosis. Last but not least, for continuous sustainable molecular diagnostics of lung cancer rational algorithms, affordable technology and appropriate reimbursement are equally necessary

Konzorcium, fő p.: Rosszindulatú daganatok ereződésének mintázata és molekuláris mechanizmusa = Consortional main: Vascularization patterns of cancer and molecular pathways behind

Vizsgálataink során a májáttétek arteriális ereződésére nyertünk bizonyítékokat és új ereződési formáját írtuk le. Pontosítottuk a daganatos intuszuszceptív érsokszorozódás celluláris mechanizmusát. Kisérletes agyi áttétekben az érinkorporációs folyamat részleteit tártuk fel, míg nyirokcsomó áttétekben a vaszkuláris invázió jelenségét igazoltuk. Bemutattuk, hogy a posztnatális vaszkulogenezishez hasonlóan daganatos betegekben a posztnatális limfatikus vaszkulogenezis is reaktiválódik prekurzorok révén, aminek prognosztikai szerepe lehet. Csontáttétes veserákokban a HIF-jelpálya EGFR-kapcsolt sajátosságait figyeltük meg. Bemutattuk, hogy az in vivo EPO kezelés képes csökkenteni kisérleti daganat HIF-jelpályájának aktivitását. Kimutattuk hogy a HIF-inaktiváló ZnSO4 kezelésnek jelentős progressziófékező hatása van melanoma modellben. Igazoltuk, hogy daganatokban az apelin új angiogén citokin. Adatokat szolgáltattunk arra hogy vastagbélrákok anti-VEGF kezelésének hatásossága a daganat K-RAS státuszától független. Bemutattuk hogy anti-VEGF kezelés EPO-val történő kombinálása hatékonyan csökkenti kisérleti daganat vérellátását. Kimutattuk, hogy VEGFR gátló kezelés hatására kisérleti daganatokban a VEGFR-független intuszuszceptív érsokszorozódás aktiválódik. | We have provided evidences for the arterial blood supply of liver metastases and defined novel vascularization forms. Studies identified details of the cellular mechanism involved in intususceptive angiogenesis of experimental tumors. We have defined details of the vessel incorporation of experimental brain metastases and provided first evidences for the existence of vascular invasion in lymph node metastases. Studies identified for the first time, that similar to postnatal vasculogenesis, lymphatic vasculogenesis is also reactivated by specific precurzors in cancer patients which might have prognostic significance. We have found in bone-metastatic renal cell cancer the further activation of the HIF-pathway associated with EGFR. We provided the first evidence that EPO administration in vivo inhibits activation of the HIF-pathway in experimental tumors. In experimental melanoma model we found that the HIF-inactivating ZnSO4 treatment has significant anti-metastatic effect. For the first time, our studies provided evidence for apelin as cancer angiogen. Clinical studies provided data on the RAS-status independent efficacy of anti-VEGF therapy of colorectal cancer. On the other hand, we have shown that combination of anti-VEGF therapy with EPO administration efficiently decreases perfusion of experimental tumors. Finaly, we have also shown that VEGFR inhibitor therapy of experimental cancer activates the VEGFR-independent intususceptive angiogenesis

Tumorspecifikus ellenanyagfragmens tisztítási és tesztelési eljárásainak kidolgozása a diagnosztikus alkalmazhatóság céljából = Development of purification and test procedures of tumorspecific immunoglobulin fragments for their further diagnostic use

A szolid tumorokban fellelhető B sejtek kiaknázása volt a cél. A tumort infiltráló B sejtek (TIL-B) tumorspecifikus antigénkötő kapacitását nemzetközileg is elsőként igazoltuk. Az eljárás diagnosztikus célú tumorfelismerő "jelölő molekulákat" eredményez. A tématervi munka sokrétű volt: 1/Új molekuláris biológiai és biotechnológiai módszerek, 2/ preparatív fehérjetisztítási munkák, 3/ összehasonlító DNS elemzések, 4/ különböző ellenanyag - antigén kötődési tesztek, 5/ immun hiszto- és citokémiai jelölési technikák történtek. Eredményeink a célkitűzéseknek megfelelőek. A medulláris emlőkarcinómából nyert egyláncú ellenanyag fragmens tisztítása sikerült. Tumorsejtkultúra lemezek és fagyasztott tumorszövet metszetek a reagensünkkel pozitív reakciót adtak immunfluoreszcencs tesztben konfokális mikroszkópos értékeléssel. Melanomák, kolorektális karcinomák és több emlőtumor típus paraffinos metszetein immunhisztokémiai eljárással is pozitív reakciót kaptunk az új GD3 gangliozid ellenanyaggal. A TIL-B immunglobulin fág megjelenítéses módszer lehetővé tette további érdekes immunglobulin variábilis régiók szelektálását invaziv ductalis emlőcarcinomákban és melanomákban. GD3 gangliozid specifikus ellenanyag fragmensünk a tumorprogresszió és invázió szempontjából fontos struktúrákat szelektíven és megfelelően jelölte a tumorszöveti mintákon. Ezen szialilált glikoszfingolipidek differenciál diagnosztikus értékét alapozta meg a munka, lehetőséget nyitva mikrometastazisok kimutatására is. | We aimed to reveal the capacity of B cells present in small amounts in solid tumors. We were internationally the first to prove the characteristic tumorspecific binding capacity of tumor infiltrating B cells (TIL-B). Our startegy helps to develop novel diagnostic tumor recognizing "labelling arrow molecules". The project work consisted of: 1/ Novel molecular biological and biotechnological techniques, 2/ preparative protein purification processes, 3/ comparative DNA analyses, 4/ different antibody ? antigen binding assays, 5/ immunohistochemical and immunocytochemical methods. The results correlated our specific aims. We could purify our medullary breast carcinoma originated antibody fragment. Tumor cell chamber slides and frozen tissue sections gave positive reactions with our reagent in immunofluorescence - confocal laser microscopy. On paraffin embedded tissue sections of melanomas, colorectal carcinomas and some other breast cancer types we got positive reaction with this unique GD3 ganglioside specific antibody. Due to our TIL-B immunglobulin phage display technique further antibody variable region genes were selected in invasive ductal breast carcinomas and melanomas. The GD3 ganglioside specific antibody labelled selectively the structures important for tumorprogression and invasion on tumor tissue specimens. The work supports the differential diagnostic value of these sialylated glycosphingolipides, opening a window towards the detection of tumor micrometastases

Differential expression of proteoglycans on the surface of human melanoma cells characterized by altered experimental metastatic potential

Heparan sulphate (HS) and chondroitin sulphate (CS) proteoglycans (PGs) frequently have opposite biologic functions in cell-matrix adhesion as well as in the regulation of cell proliferation. Data revealed that sulphated glycosaminoglycans (sGAGs) (sugar chains of PGs) are differently expressed in tumor cells characterized by different metastatic potential; the more metastatic cells contain a higher HS/CS ratio. As the proliferative capacity of tumor cells is also frequently altered in parallel with their metastatic potential, it was not clear whether observed PG alterations reflect changes in cell proliferation or metastatic potential. The cell-associated PG expression and sGAG biosynthesis was studied in tumor cells of human melanoma lines characterized by different experimental metastatic potential to the mouse liver but similar in vitro/in vivo proliferation rates. Using antibodies against PGs we found different expression of PG epitopes in melanoma lines, except from the melanoma antigen. Unlike the low CSPG (melCSPG) metastatic melanoma cells, the cell line with high metastatic capacity contained a higher proportion of positive cells for surface-HSPG without the coexpression of certain cartilage-type CSPG epitopes (recognized by MAb HSFPG 529) as well as by an increased pericellular HS/CS ratio due to intracellular accumulation/retention of CS. Immunocytochemistry of adherent cells revealed HSPGs at substrate-attached membrane areas only in cases of highly metastatic melanoma cells. These data further support our view that the absolute or relative dominance of HSPGs over CSPGs at the cell surface of metastatic tumor cells can be considered a marker of a more metastatic phenotype