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Identification of Tumor-Associated Markers for Diagnosis and Therapy

a tumor-associated marker and tumor technology, applied in the field of nucleic acids and encoded polypeptides, can solve the problem that cancer remains among the leading causes of death

Inactive Publication Date: 2014-05-15
BIONTECH AG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0023]In one embodiment, the agent is an antisense nucleic acid which hybridizes selectively with the nucleic acid coding for the tumor-associated antigen. In a further embodiment, the agent is a siRNA preferably comprising a sense RNA strand and an antisense RNA strand, wherein the sense and antisense RNA strands form an RNA duplex, and wherein the sense RNA strand comprises a nucleotide sequence substantially identical to a target sequence of about 19 to about 25 contiguous nucleotides in a nucleic acid coding for the tumor-associated antigen, preferably mRNA coding for the tumor-associated antigen. In a further embodiment, the agent is an antibody which binds selectively to the tumor-associated antigen, in particular a complement-activated or toxin conjugated antibody which binds selectively to the tumor-associated antigen. In a preferred embodiment, the antibody which binds selectively to the tumor-associated antigen is coupled to a therapeutically useful substance and / or recruits natural or artificial effector mechanisms to said cell expressing or abnormally expressing said tumor-associated antigen. In a further embodiment, the agent is a cytotoxic T lymphocyte which recognizes the tumor-associated antigen or a part thereof bound by an MHC molecule on a cell and lyses the cells labeled in this way. In a further embodiment, the agent is a T helper lymphocyte which recognizes the tumor-associated antigen or a part thereof bound by an MHC molecule on a cell and enhances effector functions of other cells specifically recognizing said tumor-associated antigen or a part thereof.
[0070]In one embodiment, the present technology relates to a pharmaceutical composition which comprises an agent that (I) inhibits expression or activity of a tumor-associated antigen and / or (II) has tumor-inhibiting activity, and is selective for cells expressing or abnormally expressing a tumor-associated antigen and / or (III) when administered, selectively increases the amount of complexes between an MHC molecule and a tumor-associated antigen or a part thereof, the tumor-associated antigen having a sequence encoded by a nucleic acid which is selected from the group consisting of: (a) a nucleic acid which comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 541, 1-540, 545, 549, 553, 557, 560, 563, 566, 570, 574, 577, 580, 583, 587, 591, 595, 599, 602, 606, 610, 613, 617, 620, and 624, a part or derivative thereof, (b) a nucleic acid which hybridizes with the nucleic acid of (a) under stringent conditions, (c) a nucleic acid which is degenerate with respect to the nucleic acid of (a) or (b), and (d) a nucleic acid which is complementary to the nucleic acid of (a), (b) or (c).

Problems solved by technology

Despite interdisciplinary approaches and exhaustive use of classical therapeutic procedures, cancers are still among the leading causes of death.

Method used

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  • Identification of Tumor-Associated Markers for Diagnosis and Therapy
  • Identification of Tumor-Associated Markers for Diagnosis and Therapy
  • Identification of Tumor-Associated Markers for Diagnosis and Therapy

Examples

Experimental program
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Effect test

example 1

Screening for Placenta-Specific Genes Aberrantly Activated in Tumors

[0222]Tissues and Cell Lines

[0223]Tissues were obtained as human surplus materials during routine diagnostic or therapeutic procedures and were stored at −80° C. until use. Cell lines were purchased from the American Type Culture Collection (ATCC) and the German Resource Collection of Microorganisms and Cell Culture (DSMZ).

[0224]RNA Isolation and Microarray Hybridization

[0225]Total RNA was isolated using the RNeasy Mini Kit protocol (Qiagen). Quantification of isolated RNA was performed using UV-spectroscopy and the quality was determined both by A260 / A280 ratio and Agilent bioanalyzer (Agilent Technologies). Five micrograms total RNA were used for cDNA synthesis with 5 pmol μl−1 T7-oligo(dT)24 primer and was performed at 43° C. for 90 minutes with the “Superscript First-Strand Synthesis-System” for RT-PCR (Invitrogen). Second-strand synthesis was performed with complete cDNA. The cDNA solution was incubated at 160°...

example 2

Validation of the Identified Tumor-Associated Markers

[0230]1. Examination of RNA Expression

[0231]The identified tumor-associated markers are first validated with the aid of RNA which is obtained from various tissues or from tissue-specific cell lines. Since the differential expression pattern of healthy tissue in comparison with tumor tissue is of decisive importance for the subsequent therapeutic application, the target genes are preferably characterized with the aid of these tissue samples.

[0232]Total RNA is isolated from native tissue samples or from tumor cell lines by standard methods of molecular biology. Said isolation may be carried out, for example, with the aid of the RNeasy Maxi kit (Qiagen, Cat. No. 75162) according to the manufacturer's instructions. This isolation method is based on the use of chaotropic reagent guanidinium isothiocyanate. Alternatively, acidic phenol can be used for isolation (Chomczynski & Sacchi, Anal. Biochem. 162: 156-159, 1987). After the tissue ...

example 3

Detailed Analysis of the Identified Tumor-Associated Markers

[0285]RNA-Isolation, RT-PCR and Real-Time RT-PCR

[0286]RNA extraction, first-strand cDNA synthesis, RT-PCR and real-time RT-PCR was performed as previously described (Koslowski, M. et al., Cancer Res. 62, 6750-6755 (2002), Koslowski, M. et al., Cancer Res. 64, 5988-5993 (2004)). Real-time quantitative expression analysis was performed in a 40 cycle RT-PCR. After normalization to HPRT (sense 5′-TGA CAC TGG CAA AAC AAT GCA-3′; antisense 5′-GGT CCT TTT CAC CAG CAA GCT-3′, 62° C. annealing) gene-specific transcripts in tumor samples were quantified relative to normal tissues using ΔΔCT calculation.

[0287]siRNA Duplexes

[0288]The SEQ ID NO:540 siRNA duplexes (Qiagen, Hilden, Germany) were directed against target sequences 5′-NNC CAC AGA AGG UAC CAG UUA-3′ (siRNA #1; sense (5′-CCA CAG AAG GUA CCA GUU AUU-3′), antisense (5′-UAA CUG GUA CCU UCU GUG GUU-3′) and 5′-NNC AGC AAG ACU CCC UCU AAA-3′ (siRNA #2; sense (5′-CAG CAA GAC UCC CUC ...

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Abstract

The present technology relates to genetic products the expression of which is associated with cancer diseases. The present technology also relates to the therapy and diagnosis of diseases in which the genetic products are expressed or aberrantly expressed, in particular cancer diseases.

Description

RELATED APPLICATIONS[0001]The present application is a continuation of International Patent Application No. PCT / EP08 / 08924, which was filed Oct. 22, 2008, claiming the benefit of priority to European Patent Application No. 07020730.3, which was filed on Oct. 23, 2007. The entire text of the aforementioned applications is incorporated herein by reference in its entirety.FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002][Not Applicable]BACKGROUND OF THE INVENTION[0003]The present technology relates to nucleic acids and encoded polypeptides which are expressed in cancers. The present technology also relates to agents which bind the polypeptides. The nucleic acids, polypeptides coded for by such nucleic acids and peptides derived therefrom, as well as related antibodies and cytolytic T lymphocytes, are useful, inter alia, in diagnostic and therapeutic contexts.[0004]Despite interdisciplinary approaches and exhaustive use of classical therapeutic procedures, cancers are still among the le...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K16/30C12Q1/68
CPCC12Q1/6886C07K16/30A61P35/00A61P37/04A61P43/00C07K14/4748C12Q2600/158C12Q2600/118
Inventor SAHIN, UGURTURECI, OZLEMKOSLOWSKI, MICHAEL
Owner BIONTECH AG
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