#line-1

Abstract:

Background: Bladder cancer is rare in young women, and advanced presentations are exceptionally uncommon. We report a de-identified case of a previously healthy 31-year-old female who developed rapidly progressive stage IV bladder cancer within 12 months of completing a three-dose Moderna mRNA vaccination series (May 2021, June 2021, December 2021).

Case Findings: Comprehensive multi-omic profiling was performed using PBIMA (Molecular Surveillance and Individualized Targeted Immunotherapy Peptide Editing) and REViSS (Spike-associated Transcriptional/Translational Instability Surveillance), incorporating analyses of plasma-derived circulating tumor DNA, whole-blood RNA, and urine exosome proteomics. Dysregulated gene expressions were identified across oncogenic driver genes (KRAS, ATM, MAPK1, NRAS, CHD4, PIK3CA, and SF3B1), auxiliary tumor-promoting signals (TOP1, PSIP1, and ERBB2), and broad evidence of genome instability with impaired DNA repair (ATM, MSH2). Within circulating tumor DNA, a host–vector chimeric read mapped to chr19:55,482,637–55,482,674 (GRCh38), in cytoband 19q13.42, positioned ~367 kb downstream of the canonical AAVS1 safe harbor and ~158 kb upstream of ZNF580 at the proximal edge of the zinc-finger (ZNF) gene cluster. This sequence aligned with perfect 20/20 bp identity to a segment (bases 5905–5924) within the Spike open reading frame (ORF) coding region (bases 3674–7480) of the Pfizer BNT162b2 DNA plasmid reference (GenBank accession OR134577.1), despite the patient only receiving Moderna vaccinations. This apparent paradox is best explained by shared Spike ORF sequences within the expression cassette across both vaccine platforms; because Moderna’s proprietary plasmid sequence has not been deposited in NCBI, BLAST defaults to Pfizer’s published reference as the nearest available match. The integration site was located outside the canonical AAVS1 “safe harbor” and within a gene-dense, recombination-prone regulatory region, raising concern for transcriptional disruption, fusion transcript formation, and oncogenic potential. The probability of a random 20-base sequence perfectly matching a predefined target is approximately 1 in a trillion, making this alignment statistically compelling and highly unlikely to be an incidental artifact.

I explored the molecular mechanisms for neurodegeneration in my last peer-reviewed paper “Melatonin regulation of phase separation in Neuro-PASC: out-maneuvering Janus-faced amyloids” [1]. As for cancer, the topic was discussed in my peer-reviewed paper on Long-COVID in 2022 [2].

A recently published peer-reviewed study that compared DNA damage in healthy, non-ICU, non-intubated ICU, and intubated-ICU COVID-19 patients, in order to determine if there is a relationship between COVID-19 infection and symptoms of Long-COVID [3].

The authors discovered that there is a distinct correlation in intensity of DNA damage with severity of COVID-19 disease. If you look at the chart below, you will see that severe infections result in higher DNA tail length and intensity [3]. What can cause these changes to DNA?

Increased DNA Tail Length and intensity -

The authors of the study employed the comet assay to assess DNA damage in COVID-19 patients. The DNA tail length refers to the distance DNA fragments migrate away from the main body of the nucleus when subjected to an electric field. It is a key parameter used to assess DNA damage, as longer tails generally indicate more extensive DNA damage. Whereas, the tail intensity refers to the proportion of DNA that migrates away from the cell's nucleus during electrophoresis, forming the "tail" in the comet-like image. Essentially, it quantifies the amount of DNA damage by measuring the fluorescence intensity of the tail relative to the overall fluorescence of the comet. A higher tail intensity indicates a greater extent of DNA damage.

The authors of the 2025 paper did not explain potential molecular mechanisms that can cause these DNA damages. But my 2022 paper presented LINE-1 derepression and hypomethylation as potential causes for DNA damage leading to genomic instability and disease [1].

Derepression of LINE-1, often observed in cancer, neurodegenerative disorders, and natural aging, can lead to increased DNA tail length and intensity due to their retrotransposition activity. This occurs because L1 elements, when not suppressed, can mobilize themselves through a process called target-primed reverse transcription (TPRT), which involves reverse transcribing an RNA copy of the L1 element into DNA and inserting it into a new genomic location. The inserted DNA often has a poly(A) tail and may be truncated at the 5' end, resulting in variable DNA tail lengths [4-6].

It is interesting to note that I am unable to find peer-reviewed reports on viruses that specifically cause LINE-1 derepression. Conversely, several peer-reviewed papers present evidence of LINE-1 derepression caused by the SARS-CoV-2 virus []7,8].

The important questions are, of course, can melatonin suppress LINE-1 derepression and attenuate potential DNA damages?

GOT MEL?” -Doris Loh

References

[1] Loh D, Reiter RJ. Melatonin regulation of phase separation in Neuro-PASC: out-maneuvering Janus-faced amyloids. Explor Neurosci. 2025;4:100678. https://doi.org/10.37349/en.2025.100678

[2] Loh, D.; Reiter, R. J. Melatonin: Regulation of Viral Phase Separation and Epitranscriptomics in Post-Acute Sequelae of COVID-19. Int. J. Mol. Sci. 2022, 23 (15), 8122. https://doi.org/10.3390/ijms23158122.

[3] Abiri, E.; Abiri, A.; Daneshi, S.; Raesi, R. The Silent Legacy of COVID-19: Exploring Genomic Instability in Long-Term COVID-19 Survivors. BMC Infect. Dis. 2025, 25, 1–9.

[4] Morrish TA, Gilbert N, Myers JS, et al. DNA repair mediated by endonuclease-independent LINE-1 retrotransposition. Nat Genet. 2002;31(2):159-165. doi:10.1038/ng898

[5] Zhang X, Zhang R, Yu J. New Understanding of the Relevant Role of LINE-1 Retrotransposition in Human Disease and Immune Modulation. Front Cell Dev Biol. 2020;8:657. Published 2020 Aug 7. doi:10.3389/fcell.2020.00657

[6] Simon M, Van Meter M, Ablaeva J, et al. LINE1 Derepression in Aged Wild-Type and SIRT6-Deficient Mice Drives Inflammation. Cell Metab. 2019;29(4):871-885.e5. doi:10.1016/j.cmet.2019.02.014

[7] Zhang, L.; Richards, A.; Barrasa, M. I.; Hughes, S. H.; Young, R. A.; Jaenisch, R. Reverse-Transcribed SARS-CoV-2 RNA Can Integrate into the Genome of Cultured Human Cells and Can Be Expressed in Patient-Derived Tissues. Proc. Natl. Acad. Sci. U. S. A. 2021, 118.

[8] Zhang, Liguo, et al. "LINE1-mediated reverse transcription and genomic integration of SARS-CoV-2 mRNA detected in virus-infected but not in viral mRNA-transfected cells." Viruses 15.3 (2023): 629.

Abstract

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