TLR3-Fc Plasmid Expression for Regulated Biomolecule Production
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Solution Overview
Problem
Bioactive molecules such as toll-like receptors, enzymes, and hormones, when over-expressed, under-expressed, or mis-expressed, lead to homeostatic imbalance and disease, necessitating therapies to regulate their production and function.
Innovation Solution
Compositions comprising recombinant plasmids encoding nucleotide sequences that upregulate the production of mRNA for fusion proteins like TLR3-Fc, TLR9-Fc, DNAse I-Fc, NGF-Fc, or insulin-Fc, administered via vectors like AAV to increase endogenous production in target cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If recombinant plasmids are used to upregulate mRNA production, then the bioavailability of target biomolecules increases, but the complexity of the treatment composition increases
Solution Approach 1:
The patent uses recombinant plasmids as intermediary carriers to deliver upregulatory sequences into target cells. These plasmids act as mediators that enable increased mRNA production without requiring direct manipulation of the target cell's genome, thus achieving the desired effect while maintaining a manageable treatment composition structure
Solution Approach 2:
The treatment employs preliminary action by introducing plasmids that contain pre-configured upregulatory sequences before the actual therapeutic effect is needed. The plasmids are prepared in advance with all necessary regulatory elements (promoters, enhancers, etc.) already in place, allowing for efficient mRNA upregulation once delivered to target cells
2Reliability
If gene expression is upregulated to treat disease, then therapeutic effect improves, but the risk of homeostatic imbalance increases
Solution Approach 1:
The patent incorporates feedback mechanisms through the use of regulated promoter sequences in the plasmid design. These promoters can respond to cellular conditions and adjust transcription rates accordingly, allowing the system to self-regulate and prevent excessive gene expression that could lead to homeostatic imbalance while maintaining effective therapeutic levels
Solution Approach 2:
The treatment utilizes parameter changes by employing inducible expression systems where gene expression levels can be precisely controlled and adjusted. By changing environmental parameters or using inducers, the system can modulate the level of biomolecule production to match therapeutic needs without causing harmful imbalances
Data Source
AI summary
Loss of B cell tolerance and generation of autoreactive anti-nuclear antibodies are hallmarks of systemic lupus erythematosus (SLE) and lupus nephritis. Lupus nephritis is characterized by glomerular and tubulointerstitial inflammation often initiated by the renal glomerular deposition of anti-nuclear immune complexes which trigger subsequent activation of complement, macrophages/monocytes and other innate inflammatory cells. The mechanism of anti-nuclear immunoglobulin accumulation and clearance in lupus nephritis pathogenesis remains largely uncharacterized. Here, we show that innate immune activation in the NZB/W F1 mouse model and in human lupus nephritis biopsies rapidly reduces DNase1 expression in renal cortex proximal tubular cells. To overcome the loss of endogenous DNase1, we treated lupus-prone mice with a hyperactive actin resistant variant of DNase1 with improved catalytic activity against nucleic acid-IgG immune complexes and acceptable in vivo pharmacokinetics. Hyperactive DNase1-Fc fusion protein ameliorates nephritis in a murine model of lupus nephritis and reduces immune complex deposition/complement fixation. Taken together, our data suggest that the loss of renal DNase1 through TLR signaling or other innate immune activation impairs clearance of autoreactive anti-nuclear immune complex deposits in the kidney to promote nephritis progression. Our findings provide a therapeutic rationale for using an engineered DNase1-Fc as a potential therapeutic approach in lupus nephritis.