28 results about "Beta thalassemia" patented technology

This invention provides vectors (e.g., lentiviral vectors) and expression cassettes for incorporation into vectors for expressing an alpha-globin (α-globin) gene. In certain embodiments method of treatment using such vectors are provided.

The invention discloses a primer group, a kit and a detection system for thalassemia gene detection. In order to more comprehensively and accurately detect point mutation, small insertion, small deletion, gene fusion mutation, large fragment deletion and gene structure variation on a thalassemia core gene and a modified gene in single detection, the invention provides a thalassemia gene detection primer group and a kit. Based on third-generation sequencing platforms such as a single-base nanopore sequencing platform and the like, all point mutation, small insertion, small deletion and gene fusion mutation in the amplification range of the thalassemia core gene and the modified gene can be detected by utilizing the primer group or the kit; and 60 alpha large fragment deletion mutations, 60 beta large fragment deletion mutations and 7 gene structure variations. The method is beneficial to rapid and comprehensive screening of thalassemia, and is high in detection sensitivity and good in accuracy.

The application discloses a pig HBB gene site-directed modification system and application. The application provides a system for site-directed modification of a pig HBB gene, the first carrier, the second carrier and the third carrier contained in the system can express gene editing proteins and sgRNA, three target sites of the HBB gene are effectively cleaved, a site-directed modification fragment of a donor DNA is used to replace a to-be-site-directed modification fragment of the target site, and accurate deletion of bases from 176 to 179 of a CDS sequence of the HBB gene is realized. 41‑42(‑CTTT) The model accurately simulates the most common beta thalassemia mutation genotype beta of Chinese population, provides a precise model for analyzing the pathogenesis of beta thalassemia of Chinese population and developing a new treatment method.

The present application relates to a primer set for detecting multiple mutations of human embryo beta-thalassemia, a kit and application thereof. The primer set comprises five primers for amplifying HBB a plurality of mutations in the gene region: HBB-F1, HBB-F2, HBB-F3, HBB-R1 and HBB-R2, the sequences of which are shown in SEQ ID NO: 1-5 respectively; and eight primers for amplifying HBB linkage SNP sites within 1Mb range of the upstream and downstream genes: L1-F, L1-R, L2-F, L2-R, L3-F, L3-R, L4-F and L4-R, the sequences of which are shown in SEQ ID NO: 6-13 respectively. Based on the specific combination of long fragment PCR amplification and long fragment high-throughput sequencing, the present application can simultaneously detect multiple mutations of beta-thalassemia in multiple embryo samples with high specificity, accuracy and speed.

Methods of using one or more fumaric acid esters or pharmacologically active salts, derivatives, analogues, or prodrugs thereof to increase expression of fetal hemoglobin (HbF) are disclosed. The methods typically include administering to a subject an effective amount of one or more fumaric acid esters optionally in combination or alternation with hydroxyurea to induce HbF expression in the subject in an effective amount to reduce one or more symptoms of a sickle cell disorder, a hemoglobinopathy, or a beta-thalassemia, or to compensate for a genetic mutation is the human beta-globin gene (HBB) or an expression control sequence thereof. Pharmaceutical dosage units and dosage regimes for use in the disclosed methods are also provided.

The application belongs to the technical field of biology, and particularly relates to a novel biomarker for screening and diagnosis of beta-thalassemia and application thereof. The application provides a novel polypeptide cluster biomarker for screening and diagnosis of beta-thalassemia based on polypeptides in plasma / serum obtained in vitro from patients, which has the advantages of high sensitivity and high specificity. The biomarker provided by the application can be detected by mass spectrometry, and the detection method is stable and efficient, and does not need to adopt cumbersome steps for inspection, thereby providing laboratory support for screening and diagnosis and treatment of beta-thalassemia.

The application belongs to the field of biology and particularly relates to a primer blocker composition, a kit, a method for detecting mutation types of beta thalassemia and application thereof. The primer blocker composition comprises a primer group and a blocker, the primer group comprises an upstream primer and a downstream primer, the upstream primer, the downstream primer and the blocker are selected from at least one of combinations I to V, and the blocker is modified with a primary amino group. The specific primer and the blocker are designed based on Gibbs free energy optimization, wild type amplification is selectively inhibited through strict matching of 3' end bases, and high specificity and high sensitivity are shown.

Provided is a method of treating or alleviating one or more pathologies associated with β- thalassemia in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising an effective amount of an anti-matriptase-2 antibody or antigen-binding fragment thereof. The one or more pathologies may be insoluble liver iron stores: α-globin precipitates in ery throid cells; reactive oxidized species formation; apoptosis of erythroid cells; reticulocytosis; or a combination thereof.

The present invention relates to products and compositions and their uses. In particular, the invention relates to nucleic acid products that interfere with or inhibit the expression of the TMPRSS6 gene in combination with one or more iron chelators and possibly other active agents, as well as to therapeutic uses such as the treatment of hemochromatosis, porphyria, and blood disorders such as beta-thalassemia, sickle cell anemia, and transfusion iron overload or myelodysplastic syndrome, as well as infections and mortality not related to relapses associated with bone marrow transplantation.

The present invention relates to the treatment of physiologic iron overload and provides antibodies against the proteolysis 2 (MTP-2). Inhibition of MTP-2 reduces dietary iron uptake and reduces iron release from cellular stores in the body. Inhibitors of MTP-2, such as antibodies against the serine protease domain, are useful in the treatment of iron overload, which is characteristic of diseases such as beta-thalassemia and which would otherwise lead to toxic accumulation of iron. Combinations of MTP-2 inhibitors with activin receptor ligand traps or with erythropoietin provide additional therapeutic effects.

The present application relates to a primer set for detecting multiple mutations of human embryo beta-thalassemia, a kit and application thereof. The primer set comprises five primers for amplifying HBB a plurality of mutations in the gene region: HBB-F1, HBB-F2, HBB-F3, HBB-R1 and HBB-R2, the sequences of which are shown in SEQ ID NO: 1-5 respectively; and eight primers for amplifying HBB linkage SNP sites within 1Mb range of the upstream and downstream genes: L1-F, L1-R, L2-F, L2-R, L3-F, L3-R, L4-F and L4-R, the sequences of which are shown in SEQ ID NO: 6-13 respectively. Based on the specific combination of long fragment PCR amplification and long fragment high-throughput sequencing, the present application can simultaneously detect multiple mutations of beta-thalassemia in multiple embryo samples with high specificity, accuracy and speed.

One aspect of the invention is a method for amplifying alpha globin genes HBA1, HBA2 and HBA12 in a single PCR tube to determine an HBA genotype of a subject. This method employs five primers selected to accurate and sensitively identify the HBA1, HBA2, and HBA12, a gene found at a higher frequency in citizens of Saudi Arabia, by accurately annealing to nucleic acids in a biological sample and simultaneously amplifying sequences encoding the alpha globin genes. This invention includes a procedure and required reagents for the amplification of alpha globin genes in a single PCR tube.

The invention belongs to the technical field of biology, and particularly relates to a novel biomarker for screening and diagnosing beta-thalassemia and application of the novel biomarker. The invention provides a novel polypeptide cluster biomarker for screening and diagnosing beta-thalassemia based on polypeptide in plasma / serum obtained in vitro from a patient, and the novel polypeptide cluster biomarker has the advantages of high sensitivity and high specificity. According to the biomarker provided by the invention, detection can be carried out through a mass spectrometry technology, the detection method is stable and efficient, tedious steps for detection are not needed, and laboratory support is provided for screening, diagnosis and treatment of beta-thalassemia.

The invention discloses a method and a composition for activating zeta-globin gene expression. The method comprises artificially forming an enhancer element comprising an NTG-N (7-8)-WGATAR sequence, an NAA-N (7-8)-WGATAR sequence, a YTATCW-N (7-8)-CAN sequence and / or a YTATCW-N (7-8)-CTN sequence on a sense strand or an antisense strand of a non-coding region of a zeta-globin gene by using a gene editing technique. The invention also discloses gRNA, ssODN, a composition and a cell, and application of the gRNA, the ssODN, the composition and the cell in preparation of a medicine for treating alpha-thalassemia. According to the method, overexpression of exogenous globin genes is not needed, and the safety risk of gene therapy is reduced. Moreover, expression of the zeta-globin gene is activated, so that the method can be suitable for various types of alpha-thalassemia patients and alpha-gene deletion or mutation patients, and is not limited to alpha-thalassemia caused by a certain mutation site.

The present invention relates to a polynucleotide construct comprising a hemoglobin subunit alpha-hemoglobin subunit beta (HBA-HBB) hybrid gene construct for enhancing the potency of gene therapy for the β-hemoglobinopathies, particularly β-thalassemia and sickle cell disease (SCD). The polynucleotide construct comprising elements that are operatively connected to each other in the following sequence: HBB locus control region (HBB-LCR), HBA promoter, and β-like globin gene. Additionally, the HBA-HBB gene construct further comprises a microRNA-based short hairpin RNA (shRNAmir) targeting one of the endogenous HBA or HBB genes to increase the ratio of vector-encoded HBBT87Q to endogenous HBA and β-like globin chains while maintaining high vector titers.

The invention further discloses application of a pharmaceutical composition containing the novel compound containing the diaza subunit sulfonyl structure in preparation of drugs for treating anemia-related diseases. The anemia-related diseases may relate to myelodysplastic syndrome (MDS), hemoglobinopathy, sickle-type cell anemia (SCD), beta-thalassemia, hereditary non-spherical cell hemolytic anemia, hemolytic anemia, hereditary spherical polycythemia, hereditary elliptical polycythemia, non-beta lipoproteinemia, paroxysmal nocturnal hemoglobinuria, and the like. Acquired hemolytic anemia or congenital anemia or chronic anemia.

The application discloses a preparation method of a dual-mode CD122 gene thalassemia detection self-powered biosensor, successfully applies an enzyme biological fuel cell (EBFC) to dual-mode detection of CD122 gene thalassemia, realizes quantitative analysis through changes of EBFC output voltage and changes of electrolyte color (RGB Blue value), and mainly concentrates on a cathode of the EBFC in the self-powered design. When CD122 exists, an electrode starts catalytic hairpin self-assembly (CHA) amplification, realizes recycling of CD122, and provides a binding site for subsequent hybridization chain reaction (HCR) amplification, at this time, methylene blue (MB) adsorbed by a biological cathode increases, and an electric signal is enhanced, and meanwhile, electrolyte color becomes lighter (RGB Blue value becomes larger). The self-powered CD122 dual-mode biosensor based on the EBFC disclosed by the application can realize simple, rapid, sensitive and efficient detection of a target.

Enhancing fetal hemoglobin (HbF) production through targeted promoter editing using base editing technologies. An embodiment includes a composition for enhancing fetal hemoglobin (HbF) expression in mammalian cells, the composition comprising a base editing system comprising at least two guide RNAs comprising targeting sequences selected from SEQ ID NO: 1 to SEQ ID NO: 5. Specifically, it involves the design and use of single guide RNAs (sgRNAs) targeting key regulatory elements within the gamma-globin (HBG1 / 2) promoter. The method combines the use of adenosine base editors to introduce mutations, creating new binding sites for transcriptional activators like TAL1 and KLF1, and disrupting repressor binding sites such as BCL11A and ZBTB7A / LRF. This dual-targeting approach significantly increases HbF expression, offering a potential therapeutic strategy for treating beta-hemoglobinopathies like sickle cell disease and beta-thalassemia.

The invention discloses a primer probe combination for beta-thalassemia single-tube multi-gene detection and a kit of beta-thalassemia single-tube multi-gene detection. According to the primer probe combination for single-tube multiple nucleic acid detection based on the nucleic acid invasive reaction and the kit thereof, provided by the invention, mutation type, insertion type and deletion type beta-thalassemia can be detected in a single closed tube at the same time through one test, the operation is simple and convenient, the pollution risk is low, and the single base recognition specificity is strong.

Antibodies against the protease 2 (MTP-2) are provided. Inhibition of MTP-2 reduces dietary iron uptake and reduces release of iron from cellular stores in the body. Inhibitors of MTP-2, such as antibodies against the serine protease domain, are useful in the treatment of iron overload, which is characteristic of diseases such as beta-thalassemia and would otherwise result in toxic accumulation of iron. Combinations of MTP-2 inhibitors with activin receptor ligand traps or with erythropoietin provide additional therapeutic effects.

Therapeutic methods, systems, and compositions for treating a patient suffering from hemoglobinopathy include treating the patient to increase expression of noncoding RNA activated by DNA damage (NORAD). Overexpression of NORAD long non-coding RNA leads to an increase in fetal hemoglobin expression. The hemoglobinopathy may be β-thalassemia or sickle cell disease.

Provided are gRNA molecules, and nucleic acids, complexes, compositions, cells, and kits thereof. Also provided are methods of increasing the expression level of HbF in progeny erythrocytes of human CD34 + HSPCs by genome editing, which are useful in the treatment of hemoglobinopathies, such as beta-thalassemia and sickle cell disease.

The invention relates to a method for constructing a long-fragment thalassemia structure variation DNA template, which comprises the following steps: respectively designing specific primers for upstream and downstream of a large fragment deletion position of a thalassemia related gene, adding uracil modified and complementarily paired connection primers, after amplification is completed, cutting by using UDG enzyme to form a viscous single-stranded terminal, and then connecting, so as to obtain the long-fragment thalassemia structure variation DNA template. Background residues are further removed through nested PCR, and finally the thalassemia structure variation long fragment DNA template is constructed. The method is simple and rapid to operate, low in cost and high in product purity, and a plurality of thalassemia structure variation long fragment DNA templates with the length larger than or equal to 15 kb and different genome complexity degrees are finally obtained by optimizing the primers and the system and can be used for evaluating the amplification performance of the primers and the like. The blank of the prior art in the aspect of efficiently constructing the specific complex long fragment template is filled, and the method has extremely high practical value and wide market application prospect.

The application relates to a primer group, a kit and a method for detecting a plurality of disease-related gene mutation types. The application can simultaneously detect genetic deafness, thalassemia and spinal muscular atrophy disease-related gene mutation types, only needs one tube, and can simultaneously detect clinically common known pathogenic SNPs, INDELs and copy number variations in one experiment, thereby significantly improving the detection efficiency of the three diseases and the standardization degree of clinical detection. The primer group comprises: 1) a primer for detecting genetic deafness gene mutation types, as shown in SEQ ID NO: 1-30; 2) a primer for detecting thalassemia gene mutation types, as shown in SEQ ID NO: 31-340; 3) a primer for detecting spinal muscular atrophy gene mutation types, as shown in SEQ ID NO: 341-350; and 4) a primer for detecting a beta-actin housekeeping gene, as shown in SEQ ID NO: 351-352.

The invention discloses an anti-human hemoglobin HBA monoclonal antibody hybridoma cell strain combination and application thereof, a purified human hemoglobin HBA subunit is used as an immunogen, after a mouse is immunized, a hybridoma is obtained by adopting an electrofusion technology, HBA and homologous subunits HBB, HBD and HBG1 are used for multi-antigen cross screening, and the HBA monoclonal antibody hybridoma cell strain combination is obtained. Two hybridoma cell strains capable of stably secreting the HBA monoclonal antibody are obtained; an antibody pair obtained by purifying the cell strain, 1A10 as a detection antibody and 19A3 as a capture antibody form a double-antibody sandwich system, and the double-antibody sandwich system can be used for ELISA quantitative detection and colloidal gold lateral chromatography test strips to realize in-vitro rapid screening of alpha-thalassemia and fecal occult blood in an in-vitro sample. The test strip, the kit and the matched detection system have the advantages of high specificity, simplicity and convenience in operation and no need of instruments, and are suitable for clinical and on-site large-scale screening.

The invention relates to the technical field of biological medicines, in particular to a recombinant lentiviral vector for treating alpha thalassemia and application of the recombinant lentiviral vector. The invention provides a lentiviral vector, a lentivirus and application of the lentiviral vector and the lentivirus in preparation of modified hematopoietic stem cells. The lentiviral vector comprises an alpha globin coding sequence and an HBB gene locus regulation sequence which are connected in series, the alpha globin coding sequence comprises a promoter, a protein coding sequence and an enhancer which are connected in series; wherein a third exon of the protein coding sequence has c.396Tgt, and a fourth exon of the protein coding sequence has c.396Tgt; c mutation; the HBB gene locus regulatory sequence comprises HS4, HS3 and HS2 which are sequentially connected in series. The provided lentiviral vector (lentiviral particle) is applied to transduction of hematopoietic stem cells, and the hematopoietic stem cells are transplanted back to the body of a patient, so that the purpose of treating alpha thalassemia can be achieved.

Provided herein, in particular, are a conjugate comprising a protein (e.g., an antibody) that specifically binds to a transferrin receptor (TFR) (e.g., human TFR (hTFR) (e.g., hTFR1)), wherein the conjugate comprises (b) a conjugate operably linked to at least one oligonucleotide that modulates the expression and / or activity of a target gene, target nucleic acid (e.g., mRNA) and / or target protein expressed by the target cell, as well as a method for producing the conjugate and a pharmaceutical composition comprising the conjugate. Furthermore, provided herein are methods of using the conjugate, including, for example, a method for treating hemoglobin disorders (e.g., sickle cell disease (SCD)) or thalassemia (e.g., α-thalassemia, β-thalassemia, δ-thalassemia, or γ-thalassemia).

The clinical history of β-hemoglobinopathies shows that the severity is mitigated by the reduction of α-globin expression, resulting from co-inheritance of α-thalassemia. The inventors identified several mutations (T>C or A>G) that can disrupt binding motifs of transcription factors using CBE- and ABE-mediated base-editing approaches. In particular, the inventors designed gRNAs that, when combined with CBEs or ABEs, disrupt binding sites for transcriptional activators (GATA1 and NF-E2) in the MCS-R2 and recapitulate the beneficial α-globin reduction observed in patients presenting both β-hemoglobinopathies and α-thalassemia. Accordingly, the present invention relates to base editing approaches for the treatment of β-hemoglobinopathies.