40 results about "LGR5" patented technology

The present invention relates to compositions and methods for characterizing, diagnosing and treating cancer. In particular, the present invention identifies LGR5 as a protein over-expressed in solid tumor stem cell. The present invention further identifies an interaction between RSPO1 and LGR5 as an alternative pathway for the activation of beta-catenin signaling. In certain embodiments, the present invention provides biomolecules that disrupt functional signaling via a LGR protein, including, in certain embodiments, molecules that inhibit the interaction between one or more RSPO proteins and one or more LGR proteins, such as LGR5. In certain embodiments, the present invention provides methods of treating cancer comprising disrupting functional LGR signaling and inhibiting growth of a solid tumor comprising solid tumor stem cells.

The present invention relates to compositions and methods for characterizing, diagnosing and treating cancer. In particular, the present invention identifies LGR5 as a protein over-expressed in solid tumor stem cell. The present invention further identifies an interaction between RSPO1 and LGR5 as an alternative pathway for the activation of beta-catenin signaling. In certain embodiments, the present invention provides biomolecules that disrupt functional signaling via a LGR protein, including, in certain embodiments, molecules that inhibit the interaction between one or more RSPO proteins and one or more LGR proteins, such as LGR5. In certain embodiments, the present invention provides methods of treating cancer comprising disrupting functional LGR signaling and inhibiting growth of a solid tumor comprising solid tumor stem cells.

The present invention relates to compositions and methods for characterizing, diagnosing and treating cancer. In particular, the present invention identifies LGR5 as a protein over-expressed in solid tumor stem cell. The present invention further identifies an interaction between RSPO1 and LGR5 as an alternative pathway for the activation of beta-catenin signaling. In certain embodiments, the present invention provides biomolecules that disrupt functional signaling via a LGR protein, including, in certain embodiments, molecules that inhibit the interaction between one or more RSPO proteins and one or more LGR proteins, such as LGR5. In certain embodiments, the present invention provides methods of treating cancer comprising disrupting functional LGR signaling and inhibiting growth of a solid tumor comprising solid tumor stem cells.

Described are cell culture solutions and systems for epithelial stem cell and organoid cultures, formation of epithelial constructs and uses of the same in transplantation. A single layer of epithelial cells that actively self-renews and is organized into crypts and villi clothes the intestine. It has been recently shown that the renewal of intestinal epithelium is driven by Lgr5+ intestinal stem cells (ISC) that reside at the base of these crypts (Barker et al., 2007). Lgr5+ stem cells can be isolated and cultured in vitro to form organoids containing crypt-vcllus structures that recapitulates the native intestinal epithelium (Sato et al., 2009).

Described are cell culture solutions and systems for epithelial stem cell and organoid cultures, formation of epithelial constructs and uses of the same in transplantation. A single layer of epithelial cells that actively self-renews and is organized into crypts and villi clothes the intestine. It has been recently shown that the renewal of intestinal epithelium is driven by Lgr5+ intestinal stem cells (ISC) that reside at the base of these crypts (Barker et al., 2007). Lgr5+ stem cells can be isolated and cultured in vitro to form organoids containing crypt-vcllus structures that recapitulates the native intestinal epithelium (Sato et al., 2009).

An objective of the present invention is to provide: a cancer stem cell isolated using a cell marker; a substantively homogeneous cancer stem cell population including said cancer stem cell; and a method of preparing said cancer stem cell population. Another objective of the present invention is to provide: a method for separating cancer stem cells with a high proliferative potential and those with a low proliferative potential; a method for inducing cancer stem cells to have a different proliferative potential; and cancer stem cells separated or induced by these separation or induction methods. A further objective of the present invention is to provide: a method of screening for pharmaceuticals using said cancer stem cell or cancer stem cell population; a method for detecting the presence of said cancer stem cell or cancer stem cell population and for identifying or quantifying the same. The present inventors discovered that highly pure colon cancer stem cells (CSC) can be obtained in a large quantity, and identified two types of states of colon CSCs distinguishable through Lgr5 expression.

Provided herein is a method for diagnosing / prognosing a metastatic cancer in a subject by measuring and detecting one or more of CS-ANXA2, DCAMKL, Lgr5 or CS-ANAX2 and DCAMKL or CS-ANXA2 and Lgr5 positive circulating tumor stem cells in the subject's blood or plasma. Also provided is a method for distinguishing the presence of early stage primary cancer from advanced stage metastatic cancer in the subject by measuring and detecting AnnexinA2, CS-ANXA2 and DCAMKL−1 or Lgr5 in the blood or plasma. In addition, there is provided a method for distinguishing the presence of benign, pre-cancerous tumorous growths or cancerous tumors in the subject by measuring and detecting AnnexinA2 and circulating tumor stem cells positive for CS-ANXA2 and DCAMKL or CS-ANXA2 and Lgr5 in the blood or plasma.

An objective of the present invention is to obtain two types of substantively homogeneous cancer stem cell populations which can be characterized using the cell surface marker Lgr5, and to provide cancer therapeutics using an antibody against a cell membrane molecule specifically expressed in these cancer stem cells by identifying said cell membrane molecule. A further objective is to provide, using an antibody against a cell membrane molecule specifically expressed in cancer stem cells, a reagent for detecting cancer stem cells, and a method for diagnosing and sorting cancer patients. The present inventors discovered that highly pure large intestine cancer stem cells (CSC) can be obtained in a large quantity, and identified the two types of conditions of large intestine CSCs distinguishable through Lgr5 expression. Moreover, the present inventors discovered that an antibody against a cell membrane molecule specifically expressed in said cancer stem cells can damage said cells.

The invention relates to the field of molecular biology and specifically discloses an immortalized rabbit small intestine epithelium cell line and a construction method thereof. The small intestines of a newly born rabbit are collected, the enzymatic digestion juice containing collagenase and neutral protease is utilized to digest, and after the digestion, a method for re-suspending centrifuging and differential adherence is adopted for purifying the pit cell cluster, so that high-purity primary pit epithelium cells can be obtained. The pit epithelium cells can participate in culture and passage. The obtained pit epithelium cells are used for performing liposome transfection expression on the SV40 big T antigenic plasmid, the passage is continued, and the immortalized rabbit small intestine epithelium cell line is finally screened. The immortalized rabbit small intestine epithelium cell line can simultaneously express intestine epithelium surface marker molecules and pit basic stem cell surface molecules. The intestine epithelium surface marker molecules include but not limited to keratin 8, E-calcitonin and ki67; and the pit basic stem cell surface molecules include but not limited to Lgr5 and sox9. The immortalized rabbit small intestine epithelium cell line can be used for establishing a rabbit small intestine epithelium in vitro model and can be used for researching a nutrition absorbing mechanism, cellular pathway excavation and pathogen in vitro culture.

The invention discloses a genome for molecular typing of medulloblastoma. The genome for molecular typing of medulloblastoma includes the following 24 genes: EPHA7 gene, OTX2 gene, ROBO1 gene, TTR gene, LGR5 gene, IGF2BP3 gene, TBR1 gene, ZFPM2 gene, TRDC gene, TRAC gene, PEX5L gene, NKD1 gene, RALYL gene, GABRA5 gene, GAD1 gene, TNC gene, KCNA1 gene, EOMES gene, MAB21L2 gene, WIF1 gene, DKK2 gene, PDLIM3 gene, IMPG2 gene, and KHDRBS2 gene. In addition, the invention also discloses an application of the genome in preparing a kit and a gene chip for molecular typing of medulloblastoma. As proved by experience, the method can accurately distinguish the WNT type, the SHH type, the Group3 type and the Group4 type of medulloblastoma, and has the advantages of objective result interpretation, high accuracy and short experiment period, and has important clinical significance in carrying out precise treatment on patients.

An objective of the present invention is to obtain two types of substantively homogeneous cancer stem cell populations which can be characterized using the cell surface marker Lgr5, and to provide cancer therapeutics using an antibody against a cell membrane molecule specifically expressed in these cancer stem cells by identifying said cell membrane molecule. A further objective of the present invention is to provide, using an antibody against a cell membrane molecule specifically expressed in cancer stem cells, a reagent for detecting cancer stem cells, and a method for diagnosing and sorting cancer patients. The present inventors discovered that highly pure large intestine cancer stem cells (CSC) can be obtained in a large quantity, and identified the two types of conditions of large intestine CSCs distinguishable through Lgr5 expression. Moreover, the present inventors discovered that an antibody against a cell membrane molecule specifically expressed in said cancer stem cells can damage said cells.

Leucine-rich repeat-containing G-protein coupled receptor 5-positive (LGR5+) intestinal cells are contacted with an inhibitor of exportin 1 (XPO1), thereby producing functionally differentiated intestinal cells. The LGR5+ cells can also be contacted with a Wnt agonist. The LGR5+ cells can also be contacted with a Notch inhibitor.

The invention discloses an application to preparation of a gastrointestinal mucosa repair protective agent by using ANKRD22 as a target. The ANKRD22 is used as the target, namely gene knock-out, geneknock-out and reduction or chemicals can reduce expression of an ANKRD22 gene. The invention provides a new target for repair of gastrointestinal mucosa light and moderate injury, namely ANKRD22 geneexpression is restrained, so that stomach tissue LGR5+ stem cells can be amplified, a stomach tissue inflammatory reaction can be alleviated, and the stomach mucosa secretion can be increased.

Provided herein are constructs of micro-aggregate multicellular, minimally polarized grafts containing Leucine-rich repeat-containing G-protein coupled Receptor (LGR) expressing cells for wound therapy applications, tissue engineering, cell therapy applications, regenerative medicine applications, medical / therapeutic applications, tissue healing applications, immune therapy applications, and tissue transplant therapy applications which preferably are associated with a delivery vector / substrate / support / scaffold for direct application.

Provided herein is a method for diagnosing / prognosing a metastatic cancer in a subject by measuring and detecting one or more of CS-ANXA2, DCAMKL, Lgr5 or CS-ANAX2 and DCAMKL or CS-ANXA2 and Lgr5 positive circulating tumor stem cells in the subject's blood or plasma. Also provided is a method for distinguishing the presence of early stage primary cancer from advanced stage metastatic cancer in the subject by measuring and detecting AnnexinA2, CS-ANXA2 and DCAMKL-1 or Lgr5 in the blood or plasma. In addition, there is provided a method for distinguishing the presence of benign, pre-cancerous tumorous growths or cancerous tumors in the subject by measuring and detecting AnnexinA2 and circulating tumor stem cells positive for CS-ANXA2 and DCAMKL or CS-ANXA2 and Lgr5 in the blood or plasma.

The invention discloses an induction method of Lgr5<+> (leucine-rich repeat-containing g-protein coupled receptor 5<+>) hepatic stem cells, and application thereof in detection. According to the induction method of the Lgr5<+> hepatic stem cells, hepatic injury is recovered through induction of the Lgr5<+> hepatic stem cells by HGF and / or Rspo1; by means of self-renewal and multifunctional differentiating capacity of endogenous stem cells of tissue and organs, the endogenous stem cells of the in-vivo tissue and organs are stimulated and activated directly by soluble molecules, so that self-healing / regeneration of the tissue and the organs is promoted. It is verified that HGF and / or Rspo1 have / has an induction effect on the humanLgr5<+> hepatic stem cells.

The invention belongs to the field of biological inner ear research, and particularly discloses a biological factor and application thereof in promoting regeneration of inner ear hair cells. The biological factor is a histone deubiquitination enzyme USP16 in a single dose state, has the activity of promoting proliferation and differentiation of inner ear stem cells into hair cells, and has an amino acid sequence as shown in SEQ NO.1. The inventor finds the space-time expression mode of USP16 in the inner ear of a mouse for the first time, subsequently finds that after the gene is knocked downin mouse hair cells, hair cell apoptosis is caused such that the normal auditory function is influenced, and finally finds that after the USP16 gene is knocked down from the inner ear Lgr5 positive inner ear stem cells, proliferation of the inner ear stem cells is promoted such that hair cells proliferate. The USP16 gene disclosed by the invention lays an important foundation for subsequent research on mouse hair cell regeneration.

Provided are compositions and methods comprising a FOXO inhibitor for increasing proliferation of Lgr5+ cochlear cells, and related methods of treating hearing loss.

Provided herein is a method for diagnosing / prognosing a metastatic cancer in a subject by measuring and detecting one or more of CS-ANXA2, DCAMKL, Lgr5 or CS-ANAX2 and DCAMKL or CS-ANXA2 and Lgr5 positive circulating tumor stem cells in the subject's blood or plasma. Also provided is a method for distinguishing the presence of early stage primary cancer from advanced stage metastatic cancer in the subject by measuring and detecting AnnexinA2, CS-ANXA2 and DCAMKL−1 or Lgr5 in the blood or plasma. In addition, there is provided a method for distinguishing the presence of benign, pre-cancerous tumorous growths or cancerous tumors in the subject by measuring and detecting AnnexinA2 and circulating tumor stem cells positive for CS-ANXA2 and DCAMKL or CS-ANXA2 and Lgr5 in the blood or plasma.

The invention relates to the technical field of biology, in particular to a method for regulating and controlling proliferation and differentiation of inner ear stem cells by Rps14 and Foxg1 and application of the method. The method comprises the following steps: firstly, regulating and controlling proliferation and differentiation of Lgr5+ inner ear stem cells by an Rps14 gene: 1, culturing OC-1 cells; 2, knocking out the Rps14 gene from the siRNA; 3, carrying out isolated culture of Lgr5+ progenitor cells and Rps14 gene knockout; and 4, carrying out the biological effect of the Rps14 gene knockout Lgr5+ progenitor cells; and secondly, detecting the influence of knockout of the Foxg1 in the Lgr5+ progenitor cells on hair cell proliferation. According to the method, an Lgr5-EGFP-CreERT2 mouse and a Foxg1-floxp mouse are hybridized, and the Foxg1 is conditionally knocked out from the Lgr5+ progenitor cells, so that the fact that the cochlea of a newborn mouse can be induced to generate extra IPCs by knocking out the Foxg1 from the Lgr5+ progenitor cells is found. Rps14 genes are knocked out through OC-1 cells by utilizing different siRNAs, and the transfection effect is detected through RT-qPCR; and after the Lgr5+ inner ear stem cells are subjected to isolated culture and transfection, proliferation and differentiation are detected, and a balling differentiation experiment on the Lgr5+ stem cells shows that knock-down of the Rps14 gene can inhibit proliferation of the stem cells and has no influence on differentiation.

Described herein are combination therapies for the treatment of cancer and other diseases. In one aspect, the methods described herein for the treatment of cancer and other diseases comprise administering a RSPO-LGR pathway inhibitor in combination with a mitotic inhibitor. The present invention relates to methods of treating cancer comprising administering to a subject a therapeutically effectiveamount of an RSPO-LGR pathway inhibitor, such as an antiRSP03 antibody or anti-LGR5 antibody. In certain embodiments, the methods further comprise administration of a mitotic inhibitor to the patient. In certain embodiments the RSPO-LGR pathway inhibitor is administered about 1 day, about 2 days, or about 3 days prior to the mitotic inhibitor.

The invention discloses a method for preventing intestinal radiation damage, wherein the intestinal stem cell genes Dclk1, Msi1, Lgr5, Bmi1 and Notch1 are up-regulated through TPA, the key signal proteins PKC-betaII and Erk1 / 2 as well as the protein p90RSK are further up-regulated, and the intestinal epithelial cell proliferation is promoted to prevent intestinal radiation damage.

The present disclosure relates to use of Rivastigmine in the preparation of an anti-radiation medicament, and belongs to the field of medical technology. The present disclosure provides use of Rivastigmine in the preparation of an anti-radiation medicament. Researches show that Rivastigmine significantly promoted the proliferation of Lgr5+ stem cells receiving 10Gy radiation (after being irradiated with 10Gy radiation for 10 d, Lgr5+ stem cells cultured in a Rivastigmine-containing culture solution had an increment 14.2 times of that of the Lgr5+ stem cells cultured in a Rivastigmine-free culture solution). Rivastigmine also significantly reduced the mortality of mice after being irradiated with 10Gy radiation (after receiving 10Gy whole body irradiation, all the mice not intraperitoneally injected Rivastigmine died within 10 d; and for the mice intraperitoneally injected Rivastigmine, the survival rate of the mice was still greater than 40% 30 d later), and thus might have good radiation-proof effect.

The invention discloses a method for combined induction of increase in endogenous Lgr5+ liver stem cells and an application of the method, and in particular a method for inducing the endogenous Lgr5+liver stem cells through combination of soluble protein molecules HGF and Rspo1. The soluble ligand can reach tissue stem cells through intravenous injection, and the soluble ligand, combined with a receptor, can activate the tissue stem cells. According to the method, the Lgr5+ liver stem cells are taken as targets. According to the method provided by the invention, on the basis of self-renewal and multifunctional differentiation capacities of various tissue and organ endogenous stem cells, the tissue and organ endogenous stem cells in a body are directly stimulated via the soluble molecules,so that self-renewal / regeneration of tissues and organs can be promoted; and from the aspects of application convenience, cost saving and safety, feasibility is enhanced.

The invention discloses application of an Lgr5 gene in protection of inner ear hair cells and promotion of support cell regeneration. The C end of the Lgr5 has a nucleotide sequence as shown in SEQ NO.1, and the 190th site of the N end of the Lgr5 has a nucleotide sequence for coding alanine. Firstly, adenoviruses of Lgr5, Lgr5-C (the part after the 823 amino acid on the Lgr5 is completely cut off) and Lgr5-190 (the 190 amino acid on the Lgr5 is changed from alanine to aspartic acid) are constructed; the protection effect of the C tail end and the N tail end of the Lgr5 gene on hair cells and the regeneration effect of inner ear stem cells are researched respectively; and finally the action mechanisms of the C tail end and the N tail end of the Lgr5 gene in an inner ear are researched respectively. Therefore, the action mechanism that the Lgr5 gene protects the hair cells and regenerates supports cells into the hair cells is determined; a new theoretical thought is provided for deafness treatment; and the Lgr5 gene replaces hearing aids and artificial cochlea to help a patient recover and rebuild the auditory function.

The invention relates to application of Rivastigmine in preparation of antiradiation drugs and belongs to the technical field of medicines. According to the application of the Rivastigmine in the preparation of the antiradiation drugs, provided by the invention, researches discover that the Rivastigmine can be used for remarkably promoting propagation of Lgr5<+> stem cells subjected to 10Gy irradiation (after 10 days of the 10Gy irradiation, the propagation amount of the Lgr5<+> stem cells cultured in a Rivastigmine-containing culture solution is 14.2 times that of the Lgr5<+> stem cells cultured in a Rivastigmine-free culture solution), the death rate of mice subjected to the 10Gy irradiation can be lowered remarkably (mice free of intraperitoneal injection of the Rivastigmine fully die in 10 days after whole-body irradiation of 10Gy, and the survival rate of mice with intraperitoneal injection of the Rivastigmine is still higher than 40% in 30 days after whole-body irradiation of 10Gy), and the Rivastigmine has a good antiradiation action.

The invention relates to the technical field of biology, in particular to a method for regulating proliferation and differentiation of inner ear stem cells by Hippo and application of the method in hair cell regeneration. The invention discloses a method for regulating proliferation and differentiation of inner ear stem cells by Hippo. The method comprises the following steps: 1, detecting the expression condition of a Hippo signal channel member in a cochlea in a wild mouse; 2, researching a regulation and control mechanism of the Hippo signal on proliferation and differentiation of the inner ear stem cells which are separated in a flow manner in an in-vitro experiment; 3, researching a coordinated regulation mechanism of Hippo and Wnt signals on the inner ear stem cells in an in-vitro experiment, and the like. By using the Lgr5-EGFP-CreERT2 tool mouse, the separation of the inner ear stem cells (Lgr5 positive cells) is accurate and a large amount of the inner ear stem cells (Lgr5 positive cells) can be realized, and a relatively good research premise can be achieved: the Lgr5 positive inner ear stem cells of the mouse are sorted. In addition, in an in-vitro experiment, after the Hippo signal channel is researched to carry out balling and differentiation experiments on the sorted inner ear stem cells, the specific action mechanism of the Hippo signal channel in regulation and control of proliferation / differentiation of the inner ear stem cells and hair cell regeneration can be deeply explored.

The invention relates to the field of inner ear stem cells, in particular to a method for regulating proliferation and differentiation of inner ear stem cells through Rassf2 and application of Rassf2 in hair cell regeneration. The regulation and control method comprises the following steps: 1, detecting expression conditions of Shh or Hippo and Rassf2 in cochlea in a wild type mouse; 2, studying the effect of Shh or Hippo and Rassf2 on proliferation and differentiation of the inner ear stem cells which are separated in a flow mode in an in-vitro experiment; 3, researching the effects of Shh or Hippo and Rassf2 on a living hair cell neomycin injury model. By using the Lgr5-EGFP-CreERT2 tool mouse, the separation of the inner ear stem cells (Lgr5 positive cells) is accurate and a large amount of the inner ear stem cells (Lgr5 positive cells) can be realized, and a relatively good research premise can be achieved: the Lgr5 positive inner ear stem cells of the mouse are sorted. Besides, after the separated inner ear stem cells are subjected to balling and differentiation experiments by researching Shh, Hippo and Rassf2 in an in-vitro experiment, the specific action mechanism of Shh, Hippo and Rassf2 in regulation and control of proliferation and differentiation of the inner ear stem cells and hair cell regeneration can be deeply explored.

The invention discloses a construction method and application of a Dkk3 gene mouse model, and belongs to the technical field of biology. The invention relates to a construction method of a Dkk3 gene mouse model. The construction method comprises the following steps: step 1, culturing OC-1 cells; knocking down the Dkk3 gene in the OC-1 cell by siRNA, and detecting the knock-down efficiency; 2, taking the cochlea of a newborn Lgr5-EGFP mouse, and digesting the cochlea into single cells; separating the Lgr5+ cells from the Lgr5-EGFP cell suspension liquid; step 3, knocking down the Dkk3 gene in the Lgr5 + cell by siRNA with the optimal knocking-down efficiency in the OC-1 cell; and 4, carrying out a cell balling test, and observing and quantifying the number and diameter of spheres to evaluate the proliferation capacity of the Lgr5 + cells.