Peak Prosperity

Is integration an anti-vax hoax?

Peak Prosperity

“Kevin McKernan discusses his discovery of DNA contamination in mRNA vaccines, exploring its implications for vaccine safety, DNA integration, and cancer risks. He also addresses the presence of SV40 sequences in vaccines, regulatory oversight failures, and the importance of ongoing research to understand the long-term effects of vaccine technologies.”

Had an expanded call with Chris Martenson on the recent B2B conference. We went into SV40, self amplifying RNAs, cancer waves, monoversities and many other topics afflicting the post COViD world.

Shortly after this we were accused of taking all of the oxygen out of the medical freedom movement with a topic that isn’t the root cause of the damage. I have been very clear in many presentations on this topic that the DNA is only one issue and if properly cleaned up will not make these vaccines safe.

Beyond that, I am not in control of which topics interest people the most nor do I think people can not walk and chew gum at the same time. Multiple concerns can be raised in parallel.

This isn’t a zero sum game.

What is important for future vaccines is that they address the DNA regulations. 10ng is an insufficient metric as the type of DNA and the type of delivery matter as much as the DNA quantity. In todays modern age of next generation sequencing, there is no excuse for not itemizing the nature of the contaminating DNA. This should be obvious after the polio vaccine contamination which occurred before Fred Sanger sequenced the first phi X174 viral genome in 1977. The PhiX genome is ~5Kb and can now be fully sequenced in an individual Oxford Nanopore (ONT) read. This platform can read over 250 bases per second on a single pore. Most ONT flow cells come with 100s-1,000s of pores sequencing in parallel. A true real time fire hose of data that costs $1000 and runs in a matter of hours.

In terms of the nature of the DNA being very material, I present to you this Gene Therapy party that just published.

This is a lentiviral vector that likely has a far higher integration rate than the Pfizer SV40 plasmid but it does show you how quickly things can go off the rails once plasmids enter the nucleus. 1000s of integrations sites per patient with 10% of the patients developing a hematologic cancer. Note, not every patients got cancer but 99% had integration events in the MECOM gene. Note, they harvest stem cells from the patient and only expose those to the vector ex-vivo, thus limiting off target bio-distribution of the vector.

I have tried to track down the plasmid used in this clinical trial as someone forwarded me an Addgene vector that looked very similar to the vector used in this trial but also contained an SV40 Enhancer. Nevertheless, I cannot find evidence of SV40 Enhancers or promoters in the sequence listed in the trial documents or supplements of this paper (welcome any correction here). Lentivirus vectors produce RNA that needs to be Reverse Transcribed and integrated. They vectors usually contain an integrase that likely drive the high integration rates seen in this study.

Why are they taking these extreme risks? cALD left untreated results in death by the time you are 10-12. It is a rare X-linked disease which implies a single genome edit in stem cells combined with bone marrow transplant would repair the broken ABCD1 gene.

Cancer may be a risk you are willing to take if you’re facing death by age 10. Not certain you want these types of risks to fend off a flu-like risk, nor do you want to gamble with integration rates far lower than this on billions of people at the same time.

In summary, we are not expecting integration rates as seen in this paper. However it is an important dataset for those wedded to the belief that any contaminating and integrated DNA in the vaccines will simply be killed by the immune system. It also pushes back on the critiques, that if the DNA contamination was a real risk, 100% of vaccinated people would have cancer right now. Clearly that isn’t the case with C19 vaccines and we have no reason to believe this contamination should result in such a binary outcome when far higher integration based approaches are utilized in cALD and only show 10% of the patients with cancer while 99% are integrated.

While there are many hypothesis of harm for the vaccine, the DNA is one marker that was never consented to during an act of fraud and it likely exhibits persistence one can forensically track to biopsies with a decentralized qPCR install base. For this reason, many of us believe it deserves much more oxygen that its currently getting and we have Chris’s Substack to thank for giving it some.

ChatGPT prompt on Integration mechanims in the Lenti-D gene therapy.

The **lentiviral vector** used to treat **cerebral adrenoleukodystrophy (cALD)**, specifically in **Lenti-D gene therapy**, integrates therapeutic genetic material into the host genome through a well-characterized mechanism. This process is essential to correct the underlying genetic defect that causes cALD, which is a mutation in the **ABCD1 gene**. Lentiviral vectors are derived from **HIV-1** and are used in gene therapy because of their ability to efficiently integrate into the genome of non-dividing and dividing cells, including hematopoietic stem cells (HSCs).

### Mechanism of Genome Integration by Lentiviral Vectors

1. **Entry into Target Cells**:

- The lentiviral vector, carrying a **corrected version of the ABCD1 gene**, first **infects** the target cells, which are usually **hematopoietic stem cells (HSCs)**. These stem cells are extracted from the patient's bone marrow or peripheral blood and then transduced ex vivo with the lentiviral vector in a laboratory setting.

- The lentiviral vector is engineered to include only the therapeutic genes and necessary elements for integration, but it lacks the genes that cause HIV to replicate or cause disease.

2. **Reverse Transcription**:

- Once the lentiviral vector enters the cell, its **RNA genome** is reverse-transcribed into **complementary DNA (cDNA)** by the viral enzyme **reverse transcriptase**. This step is crucial because lentiviral genomes are initially RNA, but they must be converted to DNA to integrate into the host genome.

3. **Nuclear Import**:

- Lentiviruses are unique in their ability to infect **non-dividing cells**, like hematopoietic stem cells, because they can transport the cDNA into the nucleus. The viral complex contains proteins that facilitate nuclear entry, even when the cell is not actively dividing (which is an advantage over other types of retroviral vectors).

4. **Integration into Host Genome**:

- The viral enzyme **integrase** is responsible for inserting the cDNA into the host genome. It binds to specific sequences at the ends of the viral cDNA and catalyzes the integration into the host cell's chromosomal DNA.

- The **integration site** is typically semi-random, meaning the lentiviral vector can integrate into many locations in the host genome, although it tends to prefer **actively transcribed regions** of the genome. However, there is no strong preference for specific genes or regulatory elements, which minimizes the risk of activating oncogenes (a concern with some other retroviral vectors).

- The **integrase** enzyme creates a staggered cut in the host DNA, allowing the viral DNA to integrate seamlessly. Once integrated, the viral DNA is treated as part of the host cell’s genome and is replicated along with the cell's own DNA when the cell divides.

5. **Expression of Therapeutic Gene**:

- After integration, the therapeutic **ABCD1 gene** under the control of a strong **internal promoter** (placed in the vector construct) begins to be **transcribed and expressed** within the host cells.

- In the case of cALD, the newly integrated and functioning **ABCD1 gene** enables the production of a normal, functional **ALDP (adrenoleukodystrophy protein)**, which restores the transport of **very long-chain fatty acids (VLCFAs)** into peroxisomes for degradation.

- The corrected cells, particularly the hematopoietic stem cells, can then give rise to differentiated cells (such as microglia in the brain) that produce functional ALDP, which can help halt the disease’s progression by restoring normal VLCFA metabolism in the central nervous system.

6. **Long-Term Therapeutic Effect**:

- Lentiviral vectors offer long-term therapeutic benefit because, once integrated, the therapeutic gene is stably passed on to all daughter cells derived from the transduced stem cell, allowing sustained expression of the corrected gene.

### Advantages of Lentiviral Vectors in Treating cALD:

- **Stable Integration**: The integration of the corrected ABCD1 gene into the host genome allows for long-term expression and correction of the underlying defect.

- **Transduction of Non-Dividing Cells**: Lentiviral vectors can infect both dividing and non-dividing cells, making them particularly useful for targeting **hematopoietic stem cells**, which may not be actively dividing at the time of transduction.

- **Reduced Risk of Insertional Mutagenesis**: While there is a risk of insertional mutagenesis (in which integration could disrupt or activate genes inappropriately), lentiviral vectors have a lower tendency to integrate near **oncogenes** compared to other retroviruses. This makes them a safer choice for gene therapy.

### Summary of Lentiviral Genome Integration for cALD Treatment:

In **Lenti-D gene therapy** for cALD, a lentiviral vector carrying a functional **ABCD1 gene** infects the patient's hematopoietic stem cells. The viral RNA is reverse-transcribed into DNA, and the viral integrase enzyme integrates this DNA into the patient’s genome. The therapeutic gene is expressed to correct the underlying metabolic defect, reducing the accumulation of very long-chain fatty acids and preventing further neurological damage. This process provides long-term, stable correction of the genetic defect responsible for cALD.