12 results about "Unfolded protein response" patented technology

The invention relates to recombinant pichia pastoris for high expression of porcine pancreatic phospholipase A2 and application of the recombinant pichia pastoris, and belongs to the technical field of genetic engineering. According to the method disclosed by the invention, a vacuole sorting receptor Vps10 is knocked out by utilizing a CRISPR-Cas9 system, a CPY sorting way between a trans-golgi apparatus and a vacuole is interrupted, and ppPLA2 degradation is reduced; then, in order to relieve unfolded protein response in the endoplasmic reticulum, molecular chaperones are introduced into the genome, and synthesis of ppPLA2 is enhanced; the expression intensity of the ppPLA2 is improved by increasing the gene copy number of the ppPLA2; and finally, high-density fermentation is performed in a 3-L bioreactor, so that high-efficiency expression of the ppPLA2 in the pichia pastoris is realized.

The present invention provides a multiomics approach, which integrate single-cell RNA-sequencing (scRNA-seq) and spatiotemporal transcriptomics (ST) offering potential for dissecting transcriptional networks and revealing cell-cell interactions involved in biomolecular pathomechanisms. The present invention also provides a multimodal approach combining high-throughput scRNA-seq and ST to elucidate XLRS-specific transcriptomic signatures in two XLRS-like models with retinal splitting phenotypes, including genetically engineered (Rs1emR209C) mice and patient-derived retinal organoids harboring the same patient-specific p.R209C mutation. Through multiomics transcriptomic analysis, the endoplasmic reticulum (ER) stress / eIF2 signaling, mTOR pathway, and the regulation of eIF4 and p70S6K pathways as chronically enriched and highly conserved disease pathways between two XLRS-like models are identified. Western blots and proteomics analysis validated the occurrence of unfolded protein responses, chronic eIF2α signaling activation, and chronic ER stress-induced apoptosis. Furthermore, therapeutic targeting of the chronic ER stress / eIF2α pathway activation synergistically enhanced the efficacy of AAV mediated RS1 gene delivery, ultimately improving bipolar cell integrity, postsynaptic transmission, disorganized retinal architecture and electrophysiological responses. Collectively, the complex transcriptomic signatures obtained from Rs1emR209C mice and patient-derived retinal organoids using the multiomics approach provide opportunities to unravel potential therapeutic targets for incurable retinal diseases, such as XLRS.

The present invention provides compounds, prodrugs, and pharmaceutically acceptable salts thereof that directly inhibit IRE-1α activity in vitro. Such compounds and prodrugs are useful for treating diseases associated with the unfolded protein response or regulated IRE1-dependent decay (RIDD) and can be used as single agents or in combination therapies.

The present invention provides methods for treating conditions, diseases or disorders associated with sex hormone imbalance and specifically with a low sex hormone level or response using NAP peptide, specifically conditions, diseases or disorders mediated via unfolded protein response.

The present invention relates to the use of a regulatory nucleic acid sequences that are able to regulate gene expression in eukaryotic cells and which are responsive to the unfolded protein response (UPR). There are disclosed regulatable introns and UPR-inducible promoters, which are able to regulate gene expression. There are also disclosed recombinant expression constructs comprising such regulatory nucleic acid sequences, whereby expression of the encoded expression product can be induced by invoking the unfolded protein response (UPR) in a eukaryotic cell containing the construct, methods of using such constructs and associated vectors, cells and suchlike.

The present invention discloses a highly active unfolded protein response element and its screening method and application. The screening method can efficiently and quickly screen out UPRE elements with high response activity. The obtained highly active UPRE2 element has been improved in both response intensity and response amplitude. Tandem connection of these highly active UPRE elements can further improve their performance. Inserting the obtained highly active UPRE2 element upstream of the core region of the promoter can achieve dynamic regulation of synthetic pathway genes, improve the yield of target products such as recombinant proteins or natural products, and has broad application prospects and important practical significance.

The invention discloses application of saikoside D as an inducer of unfolded protein reaction in prostatic cancer, and belongs to the technical field of biological medicine. The invention finds for the first time that SSD activates UPR by promoting phosphorylation of 51-site Ser in EIF2alpha, causes the failure of a cancer cell endoplasmic reticulum sensor and the emergency situation that protein synthesis is obviously inhibited, and further causes cell death and inhibition of cancer cell proliferation. According to the SSD disclosed by the invention, the expression of Phospho-eIF2 alpha (Ser51) can be obviously activated, and it is clear that UPR is a potential target spot of the SSD. The SSD provided by the invention can more effectively inhibit proliferation of prostate cancer cells by enhancing UPR in prostate cancer cells and combining with a common drug Enzaluan for prostate cancer.

The present invention relates to a Saccharomyces cerevisiae for improving the production of ovalbumin by modifying the endoplasmic reticulum, belonging to the technical field of genetic engineering. In the Saccharomyces cerevisiae CEN.PK2-1C of the present invention, the ovalbumin-encoding gene is heterologously expressed, and the INU1 signal peptide is inserted upstream of the N-terminus of ovalbumin to achieve the extracellular secretion of ovalbumin; then the fluorescence protein is used to characterize the unfolded protein response in the endoplasmic reticulum, and molecular chaperones Kar2 and PDI are co-expressed with promoters of different strengths in Saccharomyces cerevisiae to promote the folding of the endoplasmic reticulum; the opi1 gene is knocked out to increase the product secretion amount; the transcription factors INO2 and INO4 are overexpressed to expand the endoplasmic reticulum, increase the protein synthesis ability of Saccharomyces cerevisiae and promote the synthesis of ovalbumin. The yield of ovalbumin produced by the constructed Saccharomyces cerevisiae through fermentation for 72 hours reaches 6.2 mg / L.

The present technology provides a combination approach for treating multidrug resistant cancer. Multidrug resistant cancers have more mitochondrial networks than drug sensitive cancers. A first agent fragments mitochondrial networks, dissociates mitochondria from the endoplasmic reticulum, and lower the threshold for apoptosis. A second active agent induces the unfolded protein response, causing stress to the endoplasmic reticulum and limiting the ability of multidrug resistant cancer cells to grow and survive. A third active agent directly activates mitochondrial apoptosis, leading to death of the cancer cells. The active agents can be combined into a nanoparticle formulation for simultaneous delivery into multidrug resistant cancer cells. The formulation serves as a nanomedicine for treatment of multidrug resistant cancers such as multidrug resistant triple negative breast cancer.

The invention provides an application of nevermectin in tumor treatment. According to the present invention, the glucosidase II catalytic subunit GANAB can be subjected to targeted inhibition with the affinity constant KD of 0.312 [mu] M, the effect of the 3-site methoxyl demethylation product Norhistidine of the nehistidine on the GANAB is weak, and the affinity constant KD of the Norhistidine is 97.59 [mu] M; neurotetrastin can inhibit GANAB to induce endoplasmic reticulum stress and unfolded protein reaction of tumor stem cells, promote differentiation of the tumor stem cells, induce death of the tumor stem cells and play a role in inhibiting malignant tumors. In a glioblastoma stem cell model, the cytotoxic activity IC50 of the nevermectin is 7.20 + / -2.10 [mu] M, and a mouse in-situ brain tumor formed by glioblastoma stem cells can be significantly reduced after intratumoral injection, indicating that the nevermectin has good anti-tumor stem cell activity.

The present invention concerns Unfolded Protein Response (UPR) activating compounds and their use as a regenerative therapy for promoting thymic regeneration.

The invention discloses siRNA for treating UMOD gene mutant autosomal dominant hereditary renal tubular interstitial nephropathy, and belongs to the field of biomedicine. The siRNA comprises a positive-sense strand and an antisense strand, and the nucleotide sequences of the positive-sense strand and the antisense strand are SEQ ID NO.1 and SEQ ID NO.2 respectively. Compared with the prior art, the UMOD point mutation mouse model is successfully constructed, and the model completely simulates typical disease phenotypes of ADTKD-UMOD patients. In-vivo and in-vitro functional verification shows that Umod mutated mice and cell models can activate unfolded protein reaction and cell ferroptosis and apoptosis for the first time. The invention further adopts targeted UMOD mRNA for intervention by using siUMOD, and the result shows that siRNA can inhibit Umod mutant cell ferroptosis and cell apoptosis, relieve endoplasmic reticulum stress caused by mutation, improve mouse renal function and delay disease progression at the same time. According to the application, an innovative and feasible method is provided for treatment of ADTKD-UMOD, and a universal platform technology is provided for single-gene hereditary nephropathy.