Search

CN-121974997-A - Truncated variants of the LBD region of nuclear receptors and uses thereof

CN121974997ACN 121974997 ACN121974997 ACN 121974997ACN-121974997-A

Abstract

The invention relates to the technical field of biological medicines, in particular to a truncated variant of a nuclear receptor LBD region and application thereof. The invention provides truncated variants of the LBD domain of nuclear receptor, which have at least one amino acid deleted at the C-terminus. The ER, PR and GR truncated variants can induce the transcriptional activation of downstream signal channels under the condition of deleting NTD, and provide a brand-new treatment target for cancer patients such as prostate cancer, breast cancer, endometrial cancer, acute lymphoblastic leukemia, lymphoma, pancreatic cancer, lung cancer and the like.

Inventors

  • YUAN YINGJIN
  • ZHANG HAORAN

Assignees

  • 天津大学合成生物前沿研究院

Dates

Publication Date
20260505
Application Date
20241031

Claims (10)

  1. 1. A truncated variant of the LBD region of a nuclear receptor, the C-segment of which lacks at least one amino acid, said nuclear receptor being an estrogen receptor, a progestogen receptor and/or a glucocorticoid receptor.
  2. 2. The truncated variant according to claim 1, characterized in that, The wild type amino acid sequence of the LBD region of the estrogen receptor is shown as SEQ ID NO. 1; A truncated variant of the LBD region of the estrogen receptor lacks 1-297 amino acid residues at the C-terminal of the amino acid sequence shown in SEQ ID NO. 1; Preferably, the truncated variant of the LBD region of the estrogen receptor lacks 236-250 amino acid residues at the C-terminal of the amino acid sequence shown in SEQ ID NO. 1; Preferably, a truncated variant of the LBD region of the estrogen receptor lacks 236, 244, 247 or 250 amino acid residues at the C-terminus of the amino acid sequence shown in SEQ ID NO. 1; more preferably, the truncated variant of the LBD region of the estrogen receptor has the amino acid sequence shown in SEQ ID NO. 4.
  3. 3. The truncated variant according to claim 1, characterized in that, The wild type amino acid sequence of the LBD region of the progestogen receptor is shown as SEQ ID NO. 2; A truncated variant of the LBD region of the progestogen receptor lacks 1-302 amino acid residues at the C-terminal of the amino acid sequence shown in SEQ ID NO. 2; preferably, the truncated variant of the LBD region of the progestogen receptor lacks 202-216 amino acid residues at the C-terminus of the amino acid sequence shown in SEQ ID NO. 2; Preferably, a truncated variant of the LBD region of the progestogen receptor lacks 202, 210, 213 or 216 amino acid residues at the C-terminus of the amino acid sequence shown in SEQ ID NO. 2; More preferably, the truncated variant of the LBD region of the progestogen receptor has the amino acid sequence shown in SEQ ID NO. 5.
  4. 4. The truncated variant according to claim 1, characterized in that, The wild type amino acid sequence of the LBD region of the glucocorticoid receptor is shown as SEQ ID NO. 3; Truncated variants of the LBD region of the glucocorticoid receptor lack 1-278 amino acid residues at the C-terminal of the amino acid sequence shown in SEQ ID NO. 3; Preferably, the truncated variant of the LBD region of the glucocorticoid receptor lacks 201 to 215 amino acid residues at the C-terminal of the amino acid sequence shown in SEQ ID NO. 3; Preferably, a truncated variant of the LBD region of the glucocorticoid receptor lacks 201, 209, 212 or 215 amino acid residues at the C-terminus of the amino acid sequence shown in SEQ ID NO. 3; More preferably, the truncated variant of the LBD region of the glucocorticoid receptor has the amino acid sequence shown in SEQ ID NO. 6.
  5. 5. A nuclear receptor mutant comprising at least one of the truncated variants, DNA binding domains and N-terminal domains of any one of claims 1-4.
  6. 6. The nuclear receptor mutant according to claim 5, comprising, in order from the N-terminus to the C-terminus: an N-terminal domain, a DNA binding domain and a truncated variant according to any one of claims 1 to 4, Or comprising a DNA binding domain and a truncated variant according to any one of claims 1 to 4; The N-terminal domain of the estrogen receptor has an amino acid sequence shown in SEQ ID NO. 7; The DNA binding domain of the estrogen receptor has an amino acid sequence shown in SEQ ID NO. 8; The N-terminal domain of the progestogen receptor has an amino acid sequence shown in SEQ ID NO. 9; The DNA binding domain of the progestogen receptor has the amino acid sequence shown in SEQ ID NO. 10; The N-terminal domain of the glucocorticoid receptor has an amino acid sequence shown as SEQ ID NO. 11; The DNA binding domain of the glucocorticoid receptor has an amino acid sequence shown as SEQ ID NO. 12.
  7. 7. A biomaterial comprising at least one of the following I) to VI): i) A nucleic acid encoding the truncated variant of any one of claims 1 to 4 and/or the nuclear receptor mutant of any one of claims 5 to 6; II) an expression cassette comprising the nucleic acid of I); III) a plasmid vector containing the nucleic acid of I) and/or the expression cassette of II); IV) a host cell transformed or transfected with the plasmid vector of III); V) a host cell having integrated in its genome the nucleic acid molecule of I) and/or II) the expression cassette; VI), IV) or V) a product obtained by culturing said host cell.
  8. 8. Use of a truncated variant according to any one of claims 1 to 4, a nuclear receptor mutant according to claim 5 or 6 and/or a biomaterial according to claim 7 in any one of the following a) to l): a) Diagnosing or aiding in the diagnosis of cancer; b) Preparing a product for diagnosis or auxiliary diagnosis of cancer; c) Prognosis evaluation of cancer; d) Preparing a product for prognosis evaluation of cancer; e) Detecting the truncated variant and/or nuclear receptor mutant; f) Preparing a product for detecting the truncated variant and/or the nuclear receptor mutant; g) Targeting degradation, inactivation, blocking of the truncated variant and/or reducing the expression level of the truncated variant, or targeting degradation, inactivation, blocking of the nuclear receptor mutant and/or reducing the expression level of the nuclear receptor mutant; h) Preparing a product for targeting degradation, inactivation, blocking of the truncated variant and/or reducing the expression level of the truncated variant, and a product for targeting degradation, inactivation, blocking of the nuclear receptor mutant and/or reducing the expression level of the nuclear receptor mutant; i) Cancer intervention therapy or drug screening; g) Preparing a product for cancer intervention treatment or drug screening; k) Constructing a cancer model; l) preparing a product for constructing a cancer model.
  9. 9. A product, comprising: A reagent for detecting a truncated variant according to any one of claims 1 to 4, a nuclear receptor mutant according to claim 5 or 6 and/or a biological material according to claim 7; The product is a product for diagnosing or assisting in diagnosing cancer, a product for prognosis of cancer, a product for intervention treatment of cancer, a product for detecting the truncated variant according to any one of claims 1 to 4, or a product for targeting degradation, inactivation, blocking the truncated variant according to any one of claims 1 to 4 and/or reducing the expression level of the truncated variant according to any one of claims 1 to 4.
  10. 10. A method of constructing a cancer model comprising over-expressing a truncated variant according to any one of claims 1 to 4, or over-expressing a nuclear receptor mutant according to claim 5 or 6 in a cell and/or animal.

Description

Truncated variants of the LBD region of nuclear receptors and uses thereof Technical Field The invention relates to the technical field of biological medicines, in particular to a truncated variant of a nuclear receptor LBD region and application thereof. Background Nuclear receptors (Nuclear Receptors, NRs) are a family of important transcription factors within cells that can regulate the expression of target genes by binding to ligands such as hormones, thereby regulating the proliferation, differentiation and metabolic processes of the cells. Abnormal activation of nuclear receptors such as ER (estrogen receptor), PR (progestogen receptor) and GR (glucocorticoid receptor) has been widely studied in the course of occurrence and development of cancer, and has been demonstrated to be one of important causative factors of various malignant tumors. In recent years, researchers have found that in addition to ligand-dependent activation, these receptors can undergo constitutive activation in the absence of exogenous ligands, continuing to drive activation of downstream signaling pathways, leading to abnormal proliferation and survival of cancer cells. This constitutive activation mechanism has profound effects on the occurrence, resistance and prognosis of cancer. However, existing therapeutic approaches do not adequately mine these constitutively activated targets, limiting the development of novel targeted therapies. ER constitutive activation is particularly prominent in breast cancer, ovarian cancer and prostate cancer, where ER is positive. ER receptors spontaneously activate target genes, whether or not estrogen is present, resulting in sustained proliferation of cancer cells. This not only results in cancer cells that remain highly growth-competent in the absence of hormonal support, but also is closely related to the development of resistance to antiestrogens such as tamoxifen or aromatase inhibitors. Similarly, ER constitutive activation has also been reported in some ovarian and prostate cancers, and its role in tumor progression is worth intensive research. PR constitutive activation is an important downstream target for ER regulation and is related to malignant progression of endometrial cancer, breast cancer and ovarian cancer. Among these cancer types, ligand-free activation of PR can promote cell proliferation, anti-apoptosis, and cancer cell invasion capacity. In addition, constitutive activation of PR is often accompanied by reduced therapeutic efficacy, reduced sensitivity to hormonal drugs, and thus results in recurrent disease or accelerated progression. This constitutive activation places higher demands on drug development and therapeutic strategies. Constitutive activation of GR is associated with leukemia, lymphoma, lung cancer, pancreatic cancer, etc., constitutive activation of GR in cancer is not only associated with certain hematological tumors (e.g., acute lymphoblastic leukemia, hodgkin's lymphoma, etc.), but also plays an important role in a variety of solid tumors, such as lung cancer, pancreatic cancer, etc. Constitutive activation of GR generally promotes the stress survival ability of cancer cells, enabling them to evade apoptosis in adverse environments and to demonstrate resistance to chemotherapeutic drugs. This activation mechanism further enhances the malignant potential of tumors by altering the metabolism and immune response of cancer cells. Thus, how to develop effective targeted therapies for constitutive activation of GR remains a challenge in the current state of the art. Technical difficulties and unmet needs although ER, PR and GR constitutive activation has been shown to play an important role in a variety of cancers, traditional targeted hormone receptor drugs are mostly based on ligand-dependent activation mechanisms, and it is difficult to effectively inhibit ligand-free activation of receptors, since the mechanism of constitutive activation is complex and often associated with structural abnormalities of receptors or subsequent signal transduction pathway disorders. Under the current technical background, the difficulty of mining targets for constitutive activation of the nuclear receptors is high, and the molecular mechanism of the targets cannot be fully revealed by the existing detection and analysis methods, so that the development of novel anticancer targeted drugs faces bottlenecks. In addition, the existing molecular detection technology is not enough to fully analyze the constitutive activation modes of the nuclear receptors, so that the effective targeted drug design is limited, and the obvious effect of treatment is difficult to obtain. For this reason, there is a need to develop new target mining techniques and therapies to overcome the challenges of constitutive activation of receptors for cancer treatment. Disclosure of Invention In view of the above, the technical problem to be solved by the present invention is to provide truncated variants of t