A Theory of a Gene Therapy for Cancer
- A Theory of a Gene Therapy for Cancer
The proposed gene therapy for cancer harnesses the power of CRISPR technology to target mitotic and endomitotic genes in cancer cells precisely. The precision of genome engineering extends to the delivery of CRISPR to the target cells, ensuring that modifications are specifically directed and thereby minimizing impact on healthy tissues — a distinct advantage over conventional chemotherapeutics.
The strategic integration of neoantigen-directed nanobodies into viral vectors, facilitated by cutting-edge recombinant DNA technology, is what elevates the targeted delivery of CRISPR components to cancer cells. As an alternative, the utilization of CRISPR on the DNA of viral vectors, to knock in the genes responsible for the expression of neoantigen-directed nanobodies, offers a viable option for CRISPR delivery.
This method of gene therapy holds the potential to surmount challenges associated with traditional chemotherapy including the advantages of precision targeting, enduring effects, and the capacity to thwart resistance mechanisms through the modulation of mitotic genes and apoptotic signaling pathways.
The incorporation of multiplexed CRISPR technology introduces a crucial aspect of the theory, enabling the simultaneous targeting of multiple genes. The identified target genes span a spectrum that both encompasses genes pivotal in DNA repair, spindle formation, apoptosis, and endomitosis and also outlines a comprehensive strategy to disrupt cancer-related processes.
This multipronged approach enhances the therapy’s comprehensiveness and efficacy, marking a new form in the landscape of potential gene therapies for cancer. This innovative and sophisticated strategy is presented as a promising avenue in the pursuit of more precise, durable, and effective cancer treatments.
2. Targeted Genes
**DNA Repair Mechanisms**
- Description: Targeting DNA repair mechanisms to prevent resistance to treatment.
2. Associated Genes:
BRCA1 (Breast Cancer Gene 1)
BRCA2 (Breast Cancer Gene 2)
ATM (Ataxia Telangiectasia Mutated)
CHEK2 (Checkpoint Kinase 2)
RAD51 (RAD51 Recombinase)
**Mitotic Spindles**
- Description: Mitotic spindles, crucial structures in cell division, play a significant role in cancer progression.
- Associated Genes:
TPX2 (Targeting Protein for Xklp2)
KIF23 (Kinesin Family Member 23)
AURKA (Aurora Kinase A)
KIF2C (Kinesin Family Member 2C)
CENPE (Centromere Protein E)
**Apoptotic Signaling Pathway**
- Description: Enhancing apoptotic signaling is vital for promoting programmed cell death in cancer cells.
2. Associated Genes:
TP53 (Tumor Protein p53)
BCL2 (B-Cell CLL/Lymphoma 2)
CASP9 (Caspase 9)
FAS (Fas Cell Surface Death Receptor)
BAX (BCL2 Associated X, Apoptosis Regulator)
**Endomitosis**
- Description: Addressing endomitosis to prevent mitotic slippage and potential aneuploid, malignant cells.
- Associated Genes:
NDC80 (NDC80 Kinetochore Complex Component)
CCNB1 (Cyclin B1)
CDK1 (Cyclin-Dependent Kinase 1)
PLK1 (Polo-Like Kinase 1)
CDC20 (Cell Division Cycle 20)
3. Cancer
Cancer is a complex and multifaceted collection of diseases, each characterized by the uncontrolled division of cells resulting from mutations in the DNA of normal, healthy cells. The hallmark feature of cancer is the ability of these mutated cells to evade the normal mechanisms that regulate cellular growth, leading to the formation of tumors. Understanding the technical aspects of cancer involves delving into primary tumor formation, the distinction between benign and malignant tumors, pathways of metastasis through the lymphatic and circulatory systems, secondary tumor formations, the risks associated with tumors impinging on vital organs, and other critical facets of oncology.
Primary tumor formation marks the initial stage of cancer development. It begins with a single mutated cell that undergoes abnormal proliferation, forming a mass of cells known as a tumor. This process often occurs in response to genetic mutations caused by various factors, including exposure to carcinogens, genetic predisposition, replication mistake, and viral infections. As the tumor grows, it acquires the ability to invade nearby tissues and establish its presence within the host organism.
Benign and malignant tumors represent two distinct categories based on their potential for harm and invasiveness. Benign tumors are generally localized, encapsulated masses of cells that do not invade nearby tissues or spread to other parts of the body. While they may pose health risks depending on their location, benign tumors are often considered non-cancerous. In contrast, malignant tumors are characterized by their invasive nature and the potential to metastasize, making them cancerous. Metastasis is the process by which cancer cells spread from the primary tumor to other parts of the body through the lymphatic and circulatory systems.
The lymphatic and circulatory systems serve as key pathways for the dissemination of cancer cells. Cancer cells can enter the lymphatic vessels, which are responsible for transporting lymph fluid throughout the body. Once in the lymphatic system, cancer cells can travel to nearby lymph nodes, leading to the formation of secondary tumors. Similarly, cancer cells can enter the bloodstream, utilizing the circulatory system to reach distant organs and tissues where they may establish secondary tumor formations. Metastatic spread significantly amplifies the complexity and severity of cancer.
Secondary tumor formations, or metastases, represent a critical aspect of cancer progression. These distant tumors can arise in organs far from the primary tumor site, posing significant challenges to treatment and reducing the overall prognosis for the patient. The ability of cancer cells to metastasize underscores the urgency for early detection and intervention to prevent the spread of the disease.
Tumors impinging on vital organs pose serious health risks and complications. As tumors grow and infiltrate surrounding tissues, they can interfere with the normal functioning of essential organs, leading to organ failure and life-threatening consequences. The location of the tumor plays a crucial role in determining the impact on organ function, and the urgency of intervention increases when vital organs are at risk.
In addition to the fundamental aspects of cancer discussed earlier, it is essential to consider the eight hallmarks of cancer, as proposed by researchers Douglas Hanahan and Robert Weinberg. These hallmarks provide a comprehensive framework for understanding the complexities of cancer biology, offering insights into the distinctive traits that collectively drive the malignant behavior of cancer cells.
- Sustaining Proliferative Signaling: Cancer cells exhibit sustained proliferative signaling, continuously receiving signals that drive their uncontrolled growth. Aberrant activation of signaling pathways, such as growth factor pathways, contributes to this hallmark, promoting cell division and evading normal growth control mechanisms.
- Evading Growth Suppressors: Normal cells have mechanisms that inhibit their own growth, acting as a safeguard against uncontrolled proliferation. Cancer cells, however, evade these growth suppressors, often through mutations that disable the checkpoints and regulatory proteins responsible for halting cell cycle progression.
- Resisting Cell Death: One hallmark of cancer involves the ability of cancer cells to resist programmed cell death, or apoptosis. Mutations can disrupt the intricate balance between pro-survival and pro-apoptotic signals, allowing cancer cells to evade death signals and persist despite genetic damage.
- Enabling Replicative Immortality: Normal cells have a finite lifespan due to telomere shortening with each cell division. Cancer cells overcome this limitation by acquiring the ability to maintain telomeres, ensuring replicative immortality. Telomerase activation or alternative lengthening mechanisms contribute to this hallmark.
- Inducing Angiogenesis: To support their rapid growth, cancer cells induce angiogenesis, the formation of new blood vessels. This hallmark ensures a constant supply of oxygen and nutrients to the growing tumor. Pro-angiogenic factors secreted by cancer cells promote the development of blood vessels from the surrounding tissues.
- Activating Invasion and Metastasis: Invasion and metastasis are key features of malignant tumors. Cancer cells acquire the ability to invade nearby tissues and, ultimately, metastasize to distant sites. Changes in cell adhesion, degradation of extracellular matrix components, and enhanced motility contribute to this hallmark.
- Deregulating Cellular Energetics: Cancer cells often exhibit alterations in cellular energetics, favoring glycolysis even in the presence of oxygen — a phenomenon known as the Warburg effect. This shift in energy metabolism supports the biosynthetic demands of rapidly dividing cancer cells.
- Avoiding Immune Destruction: Cancer cells possess mechanisms to evade detection and destruction by the immune system. Immune checkpoints, such as PD-L1/PD-1 interactions, can be exploited by cancer cells to suppress immune responses. Additionally, tumors may develop an immunosuppressive microenvironment, hindering effective antitumor immune responses.
Integrating these eight hallmarks into an understanding of cancer enhances the complexity of the disease. The interplay between these hallmarks contributes to the adaptability and resilience of cancer cells, highlighting the importance of a multi-faceted approach in developing targeted therapies. As researchers delve deeper into the molecular intricacies of cancer, the identification and targeting of these hallmarks offer promising avenues for more effective cancer diagnosis, treatment, and prevention.
In summary, cancer is a diverse and intricate collection of diseases characterized by the uncontrolled division of cells due to DNA mutations. Understanding the technical aspects of cancer involves exploring primary tumor formation, distinguishing between benign and malignant tumors, recognizing the pathways of metastasis through the lymphatic and circulatory systems, addressing secondary tumor formations, acknowledging the dangers associated with tumors impinging on vital organs, and comprehending the eight hallmarks of cancer. As researchers and healthcare professionals continue to advance their knowledge, a comprehensive understanding of these technical aspects remains crucial for effective cancer management and improved patient outcomes.
4. Therapeutics
The field of cancer therapy has undergone a remarkable transformation, encompassing a diverse array of strategies to combat the complexities of this multifaceted group of diseases. Each therapeutic modality, from traditional interventions to cutting-edge technologies, plays a crucial role in the multidisciplinary fight against cancer.
- Surgery: Surgery stands as a fundamental pillar in cancer treatment, offering a direct and often curative approach to excise tumors. Advanced surgical techniques, including minimally invasive procedures and robotic-assisted surgery, enhance precision while minimizing the impact on surrounding healthy tissues. In conjunction with surgery, antimitotic chemotherapy often plays a pivotal role, aiming to eliminate residual cancer cells and reduce the risk of recurrence.
- Chemotherapy with Antimitotics: Chemotherapy, a systemic treatment, employs drugs to target and destroy rapidly dividing cancer cells. Antimitotics, a subclass of chemotherapeutic agents, specifically interfere with the mitotic process, preventing cells from dividing and proliferating. Agents like taxanes and vinca alkaloids disrupt microtubule dynamics, crucial for mitosis, thereby inducing cell cycle arrest and apoptosis in cancer cells. These antimitotics are particularly effective against a variety of solid tumors, including breast, ovarian, and lung cancers.
- Radiation Therapy: Radiation therapy utilizes high-energy rays to damage the DNA of cancer cells, impeding their ability to divide and grow.
- Immunotherapy: Immunotherapy harnesses the immune system to recognize and eliminate cancer cells.
- Targeted Therapies: Targeted therapies focus on specific molecular abnormalities driving cancer growth.
- Gene Therapy: Gene therapy represents an evolving frontier in cancer treatment, with applications ranging from modifying faulty genes to enhancing the body’s immune response. The theory I am developing about a gene therapy for cancer would be an antimitotic gene therapy for cancer.
- Hormone Therapy: Hormone therapy is effective against hormone-sensitive cancers, such as breast and prostate cancers.
5. Molecular Genetics
The primary components of DNA are nucleotide base pairs that bond between chromosomal backbones, and DNA replication happens in the nucleus of a cell when, during the cell cycle, in the synthesis phase, proteins copy the DNA by unwinding, matching, and then rewinding the DNA with different proteins involved in the process.
The genetic code is read by a RNA that transcribes DNA by unwinding, matching, and then rewinding DNA, like with replication, and then the transcribed mRNA is transported to a ribosome for translation into a protein.
In the process of translation, there is the the RNA that is translated by each codon into an amino acid sequence in a polypeptide protein; tRNA molecules bring amino acids to the ribosome, and the ribosome reads the mRNA codons.
Gene regulation is a phenomenon in which proteins can activate genes which then express through synthesis new proteins, and an example is a protein activating a cell cycle gene whose expressed, synthesized protein might include the construction or manipulation of microtubules in the mitotic phase of the cell cycle; gene regulation can occur at various levels, including transcriptional and post-translational.
A mutation is when a nucleotide base pair changes inadvertently, and a point mutation is when a single nucleotide is changed, such as a guanine to an thymine, whereas a chromosomal mutation includes multiple base pairs and part of the phosphate backbone of the double helix; chromosomal mutations involve changes in the structure of chromosomes, such as deletions or inversions.
Epigenetics is when an environmental factor somehow activates a section of an intron which then regulates a gene, and the concept is, as Professor Robert Sapolsky lectures in his YouTube series, a development from the Crick paradigm of molecular genetics in which genes synthesize proteins that enact on the world: In the new paradigm, of epigenetics, the world is also responsible for the regulation of genes; specific epigenetic modifications include DNA methylation and histone acetylation.
The Human Genome Project was a project that completed in the early 2000s and then finished its sequencing of the human genome about two decades later.
DNA sequencing is the identification of the sequencing of DNA and the process is intricate.
6. CRISPR
CRISPR is an acronym for clustered regularly interspaced short palindromic repeats, and the fundamental role of CRISPR in molecular biology is both as a natural phenomenon that evolved in bacteria to defend the bacteria against invading pathogens by recognizing their DNA through matching it with DNA stored from other pathogens in the CRISPR section of their DNA; and also as a genomic engineering technology with which precise gene knock in and knock out operations are possible.
The Cas9 protein the CRISPR system is the nuclease that cleaves the phosphate backbone of the double helix strands that join at bonds between nucleotides and thereby catalyzes a natural repair mechanism in either a homology directed repair for a knock in or an error prone non homologous end joining for a knock out (the error prevents the gene from functioning).
The gRNA in the CRSIPR-Cas9 technology has the role of the guide of the Cas9 protein, and the gRNA guides the protein to the correct genetic locus by using a snippet of DNA that matches the DNA of the gene to be edited to check during a traversal of a chromosome if the CRISPR system has yet arrived at the correct locus.
The steps of the CRISPR-Cas9 genome editing process requires delivery of the CRISPR system, usually through a harmless virus vector like an adeno associated virus, to a cell where the CRISPR system then enters the cell and makes its way through signal transduction pathways to the nucleus of the cell and to the chromosomes where the carried DNA that matches the correct genetic locus is then used during a traversal of the chromosomes.
CRISPR has been used in the real world to enhance the health of crops and also to treat diseases like sickle cell anemia, and current efforts are underway to expand the use of CRISPR to other diseases like cancer such as by the use of CRISPR to enhance t-cells to fight cancerous cells.
Challenges of CRISPR include the delivery of the system to the appropriate cells for editing — such as cancerous cells, though through the use of neoantigen-directed nanobodies progress at targeting cancerous cells is being made — and then off site edits once the system is on the chromosomes looking for the correct genetic locus (off-target edits are to be minimized because they impede upon the efficacy of therapeutics and engineering attempts); another difficulty is that the error prone NHEJ is only prone to errors and not always error prone, which requires reduplication of the effort to find an instance of an error.
Ethical considerations of CRISPR include the use of the technology on humans and animals, which might result in unintended side effects that could be harmful to the body.
Bibliography by Topic
- CRISPR
https://innovativegenomics.org/what-is-crispr/
2. Neo-antigen viral vectors as delivery mechanisms
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10706435/
3. Anti-mitotic chemotherapeutics:
https://www.nature.com/articles/s41422-018-0018-6
https://www.nature.com/articles/cddis2012148
4. Cancer
