25 results about "MLH1" patented technology

The present invention relates to compounds of formula (I): wherein R2, R3, R4, R5, Y1, Y2, A1, A2, A3 and A4 are each as defined herein, targeting a component MLH1 of a DNA mismatch repair (MMR) process. The invention also relates to processes for preparing these compounds, pharmaceutical compositions comprising them, and their use in the treatment of proliferative diseases, such as cancer, as well as other diseases or conditions involving MLH1 activity (e.g., triad repeat disorders). # imgabs0 #

The invention discloses a probe combination for detecting genetic susceptibility genes of prostatic cancer. The probe combination comprises at least one of sequences as shown in SEQ ID NO 1-89. The invention also discloses a kit comprising the probe combination and a method for detecting the genetic susceptibility gene of the prostatic cancer by using the kit. According to the present invention, the prostatic cancer genetic susceptibility related genes such as ATM, ATR, BRCA1, BRCA2, BRIP1, CHEK2, EPCAM, FANCA, HOXB13, MLH1, MRE11A, MSH2, MSH6, NBN, PALB2, PMS2, RAD51C, RAD51D and TP53 can be detected at one time by using the hybrid capture method; the probe provided by the invention is wide in coverage, high in sequencing depth and capable of detecting embryonic line variation of all exon regions of a target gene. All related diseases of the related genes of the genetic susceptibility of the prostatic cancer can be reported, genetic modes corresponding to the diseases are provided, and the application potential and the market value are large.

The present invention relates to compounds of formula (I) that target MLH1 and / or PMS2 proteins, which are components of the DNA mismatch repair (MMR) process: wherein R 1 , R 2 , R 3 , R 4 , R 6 and R 10 are each as defined herein. The present invention also relates to methods for preparing these compounds, pharmaceutical compositions containing them, and their use in the treatment of proliferative diseases such as cancer and other diseases or conditions involving MLH1 and / or PMS2 activity.

Disclosed herein are siRNAs and antisense oligonucleotides (ASOs) specific for an mRNA sequence of a mutS homolog 2 (MSH2) gene, PMS1 homolog 2, mismatch repair system component (PMS2) gene, mutS homolog 6 (MSH6) gene, or mutL homolog 1 (MLH1) gene. Such siRNAs and ASOs can be used in methods of inhibiting DNA mismatch repair. Also disclosed are systems and methods that combine the use of these siRNAs and ASOs with prime editing technology.

The present technology relates to methods for excluding Lynch syndrome as a possible diagnosis in patients suffering from colorectal cancers or endometrial cancers. These methods are based on detecting the methylation status of the MLH1 promoter ‘C’ region in colorectal and endometrial cancer patients using an improved and highly sensitive methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assay.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-MLH1 protein monoclonal antibody and application thereof in immunodetection. Amino acid sequences of CDR1-3 on a light chain variable region of the antibody are respectively shown as SEQ ID NO.3-5, and amino acid sequences of CDR1-3 on a heavy chain variable region of the antibody are respectively shown as SEQ ID NO.8-10. The antibody provided by the invention has strong specificity, high recognition sensitivity and good binding affinity for human MLH1 protein in cells and tissues, can accurately recognize and locate target protein in cells and tissues, greatly reduces the occurrence rate of false positive and false negative results, and can be applied to a plurality of immunodetection systems, such as human MLH1 protein, human MLH1 protein, human MLH1 protein, human MLH1 protein, human MLH1 protein and human MLH1 protein. Particularly, the method has good applicability in immunoblotting and immunohistochemical systems. Moreover, the antibody provided by the invention has cross reactivity to human and mouse homologous MLH1 proteins, and also has certain applicability and good application prospects in detection of mouse MLH1 proteins.

Methods of treating cancer or enhancing sensitivity to cancer therapies including immunotherapies by increasing tumor mutation burden. The methods use therapeutics directed against DNA polymerase proofreading and / or mismatch repair enzymes such as MLH1, MSH2, MSH6 or PMS2 are provided. The therapeutics can be used alone, in tandem or in combination with other cancer therapies, in particular with immunotherapies. Also provided are antisense therapeutics targeting DNA polymerase proofreading and / or mismatch repair enzymes.

The invention discloses a positive quality control product for detecting colorectal cancer gene large fragment deletion and a preparation method thereof, the gene contains hereditary colorectal cancer related genes MLH1 / MSH2 / MSH6 / PMS 2 / EPCAM / MUTYH / STK11, and the positive quality control product is a recombinant plasmid containing four gene fragments of a first exon of a gene to be detected, a plant sequence, a last exon of the gene and RNase P. A human genome RNase P sequence is inserted into a plasmid vector according to a fragment ratio of 2: 1, and the copy number of a to-be-detected gene is verified by adopting a fluorescent quantitative qPCR method, so that whether the obtained plasmid can be used as a positive quality control product for detection is judged, and the quality inspection of the positive quality control product is completed. The preparation method provided by the invention is simple and easy to operate and good in stability, and the positive quality control product obtained by adopting the preparation method can be used as a positive internal standard quality control material for detecting large fragment deletion of the hereditary colorectal cancer gene, so that the whole experiment process is monitored, and the accuracy of experiment and detection results is improved.

The invention relates to a capture probe group, a kit and a method for detecting hepatobiliary tumor genetic susceptibility gene polymorphism, and belongs to the technical field of biology. The nucleotide sequences of the capture probe group provided by the invention are as shown in SEQ ID NO. 1 to SEQ ID NO. 445; the hepatobiliary tumor genetic susceptibility gene comprises at least one of APC, ATM, ATR, BAP1, BRCA1, BRCA2, FANCA, MLH1, MSH2, MSH6, PALB2, PMS2 and RAD51D. The invention also provides a kit containing the capture probe group and a detection method. The method has the advantages that the coverage area is wide, the embryonic line variation of all exon areas can be detected, the designed capture probe covers the full coding area sequence of the related gene, the coverage degree of the target area reaches 100%, and the average sequencing depth reaches 100 *.

Heritable pathogenic variants in the mismatch repair (MMR) pathway, also known as Lynch Syndrome (LS), can lead to the development of colon cancer and other cancers. Following mismatch, a complex of proteins consisting of MLH1, MSH2, MSH6 and PMS2 translocate into the nucleus to signal recruitment of repair mechanisms. Flow cytometry-based, functional variant assays (FVAs), were developed to determine whether variants in these MMR repair genes and / or other related genes would augment the nuclear translocation of MLH1 and MSH2 and downstream nuclear phosphorylation of ATM and ATR in response to DNA mismatches. Each assay distinguished pathogenic variants in MMR repair genes (MLH1, MSH2, PMS2 and MSH6) from benign controls. The combination of multiple assays provided robust separation between heterozygous pathogenic variant carriers and benign controls. The ability to produce distinct molecular phenotypes by these assays suggest FVA assays of MMR pathways could be used to identify LS and associated risk of colon and other cancers and could act as an adjunct to MMR gene sequencing panels in categorizing variants.

A monoclonal hybridoma cell strain, with the deposit number of GDMCC No. 64004. The monoclonal hybridoma cell strain can stably secrete an anti-human mismatch repair protein MLH1 monoclonal antibody, which has the advantages of good specificity and high affinity. The antibody can specifically bind to a mismatch repair protein MLH1, thereby significantly improving the specificity and sensitivity of the immunological detection of the mismatch repair protein MLH1. Particularly, the antibody can specifically recognize the MLH1 protein in tumor tissues of colon cancer, gastric cancer, breast cancer and endometrial cancer, and in a human colon cancer cell line and a human cervical cancer cell line, but does not recognize the MLH1 protein in colon cancer with MLH1 deficiency and a human colon cancer cell line with MLH1 deficiency. Therefore, the antibody can be widely applied to the preparation of various immunological detection kits.

The present invention claims the use of auranofin in the preparation of tumor drugs for mismatch repair deficiency and ARID1A gene deletion, especially in the preparation of drugs for treating ARID1A. ‑ / ‑ and MLH1 ‑ / ‑ Application in oncology medicine. This invention overcomes the side effects and drug resistance limitations of auranofin. It was found that auranofin specifically binds to the TOPBP1 protein, interfering with its interaction with PHF8 and FANCJ, inhibiting the replication stress response. This finding demonstrates enhanced anti-tumor efficacy in ARID1A-deficient or MLH1-deficient tumor cells. This mechanism of action, independent of traditional TrxR inhibition, offers a new perspective for auranofin therapy and opens the door to precision medicine and new uses for established drugs.

Methods of treating a cancer comprising a mutation in tumor protein p53 (TP53) or DNA mismatch repair protein Mlhl (MLH1) comprising: determining the cancer comprises the mutation and administering an agent that decreases abundance or function of a protein synthetically lethal with TP53 or MLH1 are provided. Agents that decrease abundance or function of a protein synthetical lethal with TP53 or MLH1 for use in treating a cancer comprising a mutation in TP53 or MLH1 are also provided.

The present disclosure discloses a library construction method for detecting endometrial cancer-related gene mutations based on high-throughput sequencing, and belongs to the field of biotechnology. The method can detect the mutation types of endometrial cancer-related genes MSH2, PMS2, MLH1, MSH6 EPCAM, TP53, POLE, and PTEN in surgically removed fresh pathological tissues, formaldehyde-fixed and paraffin-embedded pathological tissues, paraffin sections, and specimens of whole blood, plasma, serum, and pleural effusion, etc. It may be used for multiple target sequences in a single tube to quickly complete the library construction. The entire library construction process only takes 3 hours, and the manual operation only needs 30 minutes. Combined with high-throughput sequencing, the platform may effectively solve the current difficulty in the detection of somatic multi-gene all-exon mutations in clinical endometrial cancer samples based on small numbers of clinical samples, and the cost is low.

Methods of treating cancer or enhancing sensitivity to cancer therapies, including immunotherapies by increasing tumor mutation burden, are provided. The methods use therapeutics directed against DNA polymerase proofreading and / or mismatch repair enzymes such as MLH1, MSH2, MSH6 or PMS2. The therapeutics can be used alone, in tandem or in combination with other cancer therapies, in particular with immunotherapies. Also provided are antisense therapeutics targeting DNA polymerase proofreading and / or mismatch repair enzymes.

The invention relates to an ovarian cancer targeted medication related gene NGS detection kit and application thereof, and the kit comprises a probe group used for detecting ovarian cancer targeted medication related gene variation conditions and corresponding reagents based on a next generation sequencing technology. By means of a hybrid capture method, variation site conditions of 12 genes (BRAF, NTRK1, NTRK2, NTRK3, RET, MLH1, MSH2, MSH6, PMS2, BRCA1, BRCA2 and KRAS) related to important targeted medication of ovarian cancer can be detected at a time. The probe used in the invention has the advantages of wide coverage, high sequencing depth and strong specificity to ovarian cancer patients, and compared with polygene solid tumor large panel detection, the resource waste caused by irrelevant gene detection is reduced, and the reduction of the detection cost and the improvement of the detection efficiency are realized.

The present invention relates to a novel polypeptide that binds to MLH1 to inhibit its interaction with PMS2 and the formation of the MutLα complex, thereby enhancing prime editing efficiency, and uses thereof for prime editing. The polypeptide according to the present invention binds to MLH1 protein to inhibit the formation of a complex with PMS2, resulting in a significant improvement in prime editing efficiency while exhibiting remarkably low off-target editing and cellular toxicity. Furthermore, with a significantly smaller size compared to conventional dominant-negative MLH1 (MLH1dn), the polypeptide of the present invention can be easily integrated into various existing prime editing systems and one vector and is universally applicable. Therefore, the novel MLH1-binding polypeptide of the present invention and the nucleic acid encoding same can be advantageously used in the field of gene editing.

The invention relates to the technical field of synthetic biology and metabolic engineering. The invention provides a saccharomyces cerevisiae recombinant strain capable of improving vanillin tolerance and a construction method thereof, which are characterized in that related genes MSH1, MSH2, MSH3, MSH4, MSH5, MSH6, MLH1, MLH2, MLH3 or PMS1 of a strain DNA mismatch repair system are innovatively knocked out, so that the adaptive evolution ability of the strain is enhanced, and the limitation of traditional metabolic engineering modification is broken through; a stepped vanillin concentration domestication strategy is utilized, and a mutant strain with remarkably improved tolerance is obtained through screening. The limitation of vanillin toxicity on thallus growth and product synthesis is effectively relieved, and a certain foundation is laid for constructing an efficient vanillin biosynthesis system.

The invention provides a composition and a kit for detecting combined methylation of CpG sites in an MLH1 gene target region in human tumor tissues and application of the composition and the kit. The CpG locus combination in the MLH1 gene target area in the human tumor tissue provided by the invention comprises 10 CpG loci which have the most significant difference with methylation states in sporadic and Lingchi syndrome related tumors in the MLH1 gene. The detection composition is designed on the basis of 10 CpG loci, competitive Blocker oligonucleotides are introduced into the detection composition and can be specifically combined with a conversion sequence corresponding to non-methylated DNA, non-specific amplification of the detection composition is effectively inhibited in the early stage of PCR amplification, the common problem of'tail raising 'or background rising in conventional qPCR is thoroughly solved, and the detection composition has the advantages of high specificity, high sensitivity and the like. It is ensured that amplification signals are completely derived from target methylation alleles, the detection specificity is close to 100%, and extremely high confidence is provided for clinical diagnosis.

The present invention relates to compounds of Formula (I) that target the MLH1 and / or PMS2 proteins that are components of the DNA Mismatch Repair (MMR) process:wherein R1, R2, R3, R4, R6 and R10 are each as defined herein. The present invention also relates to processes for the preparation of these compounds, to pharmaceutical compositions comprising them, and to their use in the treatment of proliferative disorders, such as cancer, as well as other diseases or conditions in which MLH1 and / or PMS2 activity is implicated.

The invention discloses a construction method and application of an ovarian cancer BRCA1 mutant cell model based on CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / associated protein 9). According to the invention, an ovarian cancer BRCA1 (c.5091 Tgt; A and c.5521 del) endogenous point mutation cell model is constructed by adopting a CRISPR / Cas9 technology, and experimental conditions of editing BRCA1 by CRISPR / Cas9 in ovarian cancer cells are optimized by adopting multiple transfections or by interfering a mismatch repair pathway key factor MLH1, so that the mutation rate is increased. According to the invention, the novel mutation c.5091 Tgt of BRCA1 is jointly verified in a manner of combining endogenous mutation and exogenous mutation; a and c.5521del can increase the sensitivity of ovarian cancer cells to carboplatin and interfere with the expression of carboplatin sensitive genes. Theoretical and experimental basis is provided for precise diagnosis and treatment and prognosis evaluation of ovarian cancer, and more clinical problems can be solved.

This invention relates to the field of early cancer diagnostic reagents, specifically to a non-invasive intelligent diagnostic kit for early colorectal cancer based on peripheral blood and its application. The kit targets key gene mutation sites (MLH1 c.113del A, MSH2 c.788del A, MUTYH c.1005 G>C, PMS2 c.288 C>T, and c.780 C>G) screened in the Chinese population and includes a specific primer set, circulating tumor DNA (ctDNA) extraction, and sequencing library preparation reagents. The accompanying detection system analyzes mutation data using a logistic regression model trained on the aforementioned sites to achieve early screening. Clinical trials have shown that this method has good performance in early colorectal cancer screening in the Chinese population.

An embodiment of the present invention discloses a method for detecting NF1 gene mutations by multi-gene combination, belonging to the technical field of nucleic acid detection. The method includes: LR-PCR detection of the NF1 gene; multiplex PCR detection of the SPRED1, GNAS, PTPN11, MLH1, MSH2, MSH6, and PMS2 genes. The SPRED1, GNAS, PTPN11, MLH1, MSH2, MSH6, and PMS2 genes are added to the NF1 gene detection, and the detection of these genes helps to clarify the diagnosis and differential diagnosis of diseases such as Legius syndrome, Noonan syndrome, and structural mismatch repair deficiency syndrome. The LR-PCR detection of the NF1 gene achieves full-length coverage of the NF1 gene and covers the breakpoints of two types of NF1 whole-gene microdeletions, that is, the most common type is type I deletion of 1.4 Mb; the multiplex PCR detection of the SPRED1, GNAS, PTPN11, MLH1, MSH2, MSH6, and PMS2 genes covers the CDS regions of the SPRED1, GNAS, PTPN11, MLH1, MSH2, MSH6, and PMS2 genes and the pathogenic or likely pathogenic sites of these genes located in the non-coding region included in the ClinVar and HGMD databases.

The invention discloses a human MLH1 methylation detection reagent based on digital PCR (Polymerase Chain Reaction). The reagent comprises a methylation primer probe combination of MLH1, and the combination comprises an MLH1 upstream primer with a sequence as shown in SEQ ID NO.1, an MLH1 downstream primer with a sequence as shown in SEQ ID NO.2 and an MLH1 probe with a sequence as shown in SEQ ID NO.3. The invention further discloses a kit for detecting the methylation of the human MLH1 based on the digital PCR. The invention further discloses a human MLH1 methylation detection kit and a human MLH1 methylation detection method based on digital PCR. The reagent, the kit and the detection method are high in specificity and good in accuracy.

The invention discloses a composition for predicting the curative effect of gastric cancer chemotherapy based on the methylation level of a CpG site of an MLH1 gene and application of the composition, the CpG site is derived from the MLH1 gene, and the CpG site is any one of chr3: 37033633, chr3: 37033626, chr3: 37033601 and chr3: 37033490 or a combination of more than two of the chr3: 37033633, the chr3: 37033626, the chr3: 37033601 and the chr3: 37033490. According to the CpG loci in the MLH1 gene, the composition of the CpG loci, the kit, the system, the computer equipment and the computer readable storage medium provided by the invention, the methylation level of the CpG loci of the MLH1 gene in a biological sample is measured, so that the chemotherapy curative effect of the gastric cancer can be accurately predicted; and a new thought and a new method are provided for precise medical treatment of patients with gastric cancer clinically.