Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...
Reexamination Certificate
1999-09-21
2002-06-04
Caputa, Anthony C. (Department: 1642)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Blood proteins or globulins, e.g., proteoglycans, platelet...
C530S350000, C530S380000, C530S385000, C530S386000, C530S387100, C530S387700, C530S388100, C530S388200, C530S389100, C530S389700, C435S004000, C435S006120, C435S007100, C435S007210, C435S007230, C435S007900, C435S007920, C435S007940, C436S063000, C436S064000, C436S512000
Reexamination Certificate
active
06399748
ABSTRACT:
The present invention relates to an in-vitro method for prognosticating the illness development of patients with carcinoma of the breast and/or for diagnosing carcinoma of the breast, to kits suitable for performing said method, and to the use of T1-specific antibodies or fragments thereof or of T1-specific oligonucleotides for determining T1 protein or T1-mRNA in patients with carcinoma of the breast.
Carcinomas of the breast, in particular invasive carcinomas of the breast, are malign tumors with an extremely different clinical development that could so far not be predicted. Invasive carcinoma of the breast is the most frequently observed malign tumor in women; on the statistical average, every 16th woman suffers therefrom. In the case of an existing carcinoma of the breast, the breast tumor is first of all removed as the primary therapeutic measure. Especially the nodal state, but also tumor size, histological type, degree of differentiation and hormone receptor condition are nowadays regarded as important parameters for prognosis and further therapeutic planning. Emphasis is however placed on the evaluation of the state of the axillary lymph node. For instance with patients suffering from lymph node disease in the axillary region (nodal-positive, N(+)) already at the time of tumor diagnosis or surgery, a chemotherapy is normally carried out immediately, optionally supported by an additional radiation therapy. In general, patients with a negative axillary lymph node state (nodal-negative, N(0)) have a better chance and, therefore, are in general not subjected to a chemotherapeutic or radiation-therapeutic aftertreatment. Statistically, however, up to 30% of the patients classified as N(0) suffer from a relapse (Yan 1992). Such a high rate of relapse demonstrates that the factors of prognosis that have so far been known describe the illness development in a very incomplete manner only.
In recent years a great number of molecules were tested as to their possible use as prognosis factors for carcinoma of the breast (for the purpose of survey: Schmitt et al., 1994; Hoskins and Weber 1995), e.g. (proto)oncogenes such as c-erbB-2 (Allred et al., 1992, Archer et al., 1995), the tumor suppressor gene p53 (Barnes et al., 1993, Lipponen et al., 1993), the urokinase-type plasminogen activator uPA (Jaenicke et al., 1993; Wilhelm et al., 1994), the adhesion molecule E-cadherin (Rasbridge et al., 1993; Graff et al., 1995) and the cytoskeleton protein vimentin (Sommers et al., 1992). The vimentin synthesis correlates with the invasive growth of breast-carcinoma cell lines in vitro (Thompson et al., 1991) and partly with rapidly growing invasive ductal breast carcinomas having a poor prognosis in vivo (Domagala et al., 1990). Furthermore, possible factors of prognosis are formed by cytometrically determined morphometric features and texture features and also by DNA parameters (Auer et al., 1994).
However, none of the above-mentioned factors permits a sufficiently reliable prognosis of the further illness development after removal of the breast carcinoma.
It is therefore the object of the present invention to provide a method which allows for a sufficiently reliable prognosis of the further illness development in patients with carcinoma of the breast.
According to the invention this object is achieved by an in-vitro method for prognosticating the illness development of patients with carcinoma of the breast and/or for diagnosing carcinoma of the breast, which method comprises the qualitative or quantitative determination of T1 protein and/or T1-mRNA in sample material obtained from patients.
The T1 protein is an extracellular, soluble glycoprotein of 60-70 kDa (Werenskiold, 1992) with homology to members of the immunoglobulin superfamily, in particular the carcinoma-embryonal antigen (Klemenz et al., 1989).
The molecule was identified in an analysis of the former effects of oncoproteins (p21
ras
and p39
v-mos
) on the gene expression in fibroblasts (Werenskiold et al., 1989). In the mouse there is a synthesis of the T1 protein in embryonal tissues only; it cannot be detected in the adult animal (Rö&bgr;ler et al., 1995 a,b). The function of the T1 protein has not been completely darified yet, but the isolation of a second membrane-bound variant of the molecule (T1-M) points to a function as a cytokine receptor. The membrane-bound T1-M protein is very similar to the IL-1 receptor type 1, but has no affinity to the cytokines IL-1 &agr; and &bgr; (Rö&bgr;ler et al, 1995 b; Danescu and Werenskiold, 1995) or IL-1 ra (Gayle et al., 1996). T1-M is a novel mast cell-specific cytokine receptor (Rö&bgr;ler et al., 1995 b; Thomassen et al., 1995). The oncogene-inducible soluble variant of the T1 protein is a shortened form of said receptor and corresponds to the ligand-binding domain thereof. Recombinantly produced, soluble T1 from the mouse (Rupp et al., 1995) blocks the growth of mast cells.
So far, in breast carcimonas of the mouse, an overexpression of the soluble T1 protein has exclusively been observed in invasively growing, poorly differentiated tumors. Both the tumor stroma and the anaplastic tumor cells synthetize T1. In-situ hybridizations demonstrate an increased expression of T1 in tumor cells on the periphery of tumor cell complexes and, possibly induced by the tumor cells, in the stroma cells directly adjacent thereto (Rö&bgr;ler et al, 1993). The induction of the T1 expression in the tumor cells correlates with the phenotypic transformation thereof and is accompanied by a loss in the E-cadherin and cytokeratin production. It is controlled by an AP1-dependent promoter of the T1 gene which is only active in non-hemopoietic (e.g. fibroblastic) cells (Thomassen et al., 1995). During phenotypic transformation of the epithelial tumor cells the induction of T1 is distinctly performed prior to that of the also AP1-dependent, mesenchymal cytoskeleton protein vimentin and therefore forms an early marker for the transformation process.
Surprisingly enough, it has now been found that the presence or absence of a T1 transcription or expression provides information about the future illness development, in particular information about the probability of the occurrence of a relapse or of the development or growth of metastases. As will be explained in detail in the examples, a high T1 value surprisingly correlates with a positive prognosis in patients with an N(0) nodal state whereas a high T1 protein or T1-mRNA level means a negative prognosis in patients with an N(+) nodal state.
In a preferred embodiment the T1 protein and/or the T1-mRNA is determined in a tumor tissue sample of the patient. To this end tissue sections are e.g. made and fixed in accordance with standard methods and are subsequently subjected either to an immunoassay for detecting an existing T1 protein or to hybridization with oligonucleotides, which are or can be labeled, or with DNA fragments. Total RNA or poly(A)
+
-mRNA can optionally be isolated from the tumor tissue according to standard methods and, for example after a gel-electrophoretic separation or after fixation to a solid matrix, the total RNA or poly(A)
+
-mRNA can then be determined again by hybridization with an oligonucleotide which is or can be labeled.
T1 protein can be detected not only in the tumor tissue itself, but also in various body fluids of the patients afflicted. In a preferred embodiment the method of the invention is carried out with a blood or serum sample:
There are a number of methods for determining T1-mRNA. As has already been pointed out above, it is possible, on the one hand, to detect T1-mRNA in situ or in corresponding RNA or mRNA preparations by hybridization with a corresponding oligonucleotide probe or a DNA fragment. The oligonucleotide probe or the DNA fragment itself can produce a measurable signal, i.e. it may be radioactively labeled, or can be capable of producing a signal by interacting with other molecules. For an improved evaluation of the corresponding tests the signal should be amplified in most cases. The ta
Akin Gump Strauss Hauer & Feld L.L.P.
Caputa Anthony C.
GSF-Forschungszentrum fur Umwelt und Gesundheit GmbH
Harris Alana M.
LandOfFree
In-vitro method for prognosticating the illness development... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with In-vitro method for prognosticating the illness development..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and In-vitro method for prognosticating the illness development... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2979921