Modified DNase 1 Gene Construct for Targeted Cancer Cell Apoptosis
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Solution Overview
Problem
Current cancer treatments, particularly for malignant melanoma, face challenges in effectively triggering apoptosis in cancer cells due to the natural protection mechanisms of cancer cells, which include inactivating or underexpressing death receptors and overexpressing anti-apoptotic proteins, making it difficult for existing therapies to induce programmed cell death efficiently.
Innovation Solution
A gene construct is developed with a nuclear localization signal, a deleted signal peptide, an inhibitor-resistant binding site, and a promoter for exclusive expression in selected cells, allowing for targeted and efficient induction of programmed cell death without triggering the apoptosis cascade, using a modified nuclease encoding gene like DNase 1.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If native DNase 1 is used, then the protein is produced with normal structure and function, but it cannot trigger intracellular apoptosis due to lack of nuclear localization signal and presence of signal peptide
Solution Approach 1:
The signal peptide sequence is removed from the DNase 1 protein to prevent its diversion to the endoplasmic reticulum, allowing the protein to remain in the cytoplasm and nucleus where it can trigger apoptosis
Solution Approach 2:
A nuclear localization signal is added to the DNase 1 protein before apoptosis is needed, enabling the protein to access the nucleus and trigger apoptosis without requiring the apoptosis cascade to occur first
2Reliability
If DNase 1 is delivered to cancer cells, then apoptosis can be triggered, but cancer cells protect themselves through multiple mechanisms including nuclear membrane barrier and actin inhibition
Solution Approach 1:
The actin binding site of DNase 1 is mutated to create resistance against actin inhibition, preemptively counteracting one of the main protection mechanisms cancer cells use to prevent apoptosis
Solution Approach 2:
The nuclear localization signal is added before treatment to ensure DNase 1 can access the nucleus immediately upon delivery, bypassing the need for nuclear membrane disruption that normally protects cancer cells
3Adaptability or versatility
If broad-spectrum chemotherapy is used, then many cancer types can be treated, but treatment resistance develops and side effects increase
Solution Approach 1:
The DNase 1 protein is modified with specific sequences (nuclear localization signal, mutated actin binding site) that give it specialized functionality to overcome specific cancer cell protection mechanisms, making the treatment more effective against resistant cancers
Solution Approach 2:
The protein's functional parameters are changed through mutation of the actin binding site and addition of nuclear localization signal, transforming it from a protein that cannot trigger apoptosis to one that can effectively induce cell death in resistant cancer cells
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The gene construct effectively initiates and executes programmed cell death in cancer cells, overcoming the protective mechanisms of cancer cells and achieving high killing efficiency with reduced side effects by directly targeting and destroying cancer cells without affecting healthy cells.
Implementation Method 1
adding a nuclear localization signal
Implementation Method 2
DNase 1, for example, is a powerful DNA-degrading enzyme, that preferentially cleaves DNA at phosphodiester linkages adjacent to a pyrimidine nucleotide
Implementation Method 3
The signal peptide, which diverts the protein away from the nucleus, was not removed
Implementation Method 4
the DNase 1 protein is equipped with a signal peptide (SP), which directs it away from the nucleus, to the endoplasmic reticulum, where it is packaged in an endosome to contain its activity
Data Source
AI summary
A gene construct comprising a programmed-cell-death executioner gene having a nuclear localization signal, a deleted signal peptide, an inhibitor-resistant binding site, a promoter, and an activator. A method of making a gene construct, by modifying a programmed-cell-death executioner gene by adding a nuclear localization signal, deleting a signal peptide, mutating a binding site for an inhibitor to make it inhibitor-resistant, adding a promoter for exclusive expression in selected cells, and adding an activator. A method of eliminating undesired cells from a patient. A method of treating cancer. An array comprising at least two gene constructs wherein all of the gene constructs differ with respect to the programmed-cell-death executioner gene and the nuclear localization signal. A method of personalizing anti-cancer treatment. A method of increasing DNase 1 resistance to actin binding. A method of increasing catalytic activity of DNase 1 binding.