Breast cancer metastasis, NNMT, SIRT1, microRNA449a; yuanhadine and pomegranate peel extract may be protective.

The short story in my opinion is: keep your one carbon methylation cycles spinning normally so that you don't have excess NNMT issues.

Pomegranate peel extract is consistently good at preventing cancer cell metastasis and a recent research finding may be a clue as to why.

NNMT is an enzyme involved in the one carbon methylation cycles which I have mentioned are a factor in cancer risk. Without one carbon methylation cycle function, cancer and other chronic conditions related to DNA changes may occur, like Parkinson's disease, linked to mitochondrial DNA changes.

Functional methylation cycles are needed to methylate DNA which keeps some of it turned off instead of being in a constant growth mode.

Overexpression of NNMT is seen in cancer cells - I would think the cell is trying to get that methylation cycle rolling again.

• See this post for that background info: (9) Nature loves a good design; Cancer, POTS, Epigenetics & the One-Carbon Methylation Cycles. (substack.com)

What happens instead is the extra NNMT can promote collagen formation which would help a free cancer cell to take hold somewhere else and start growing there too.

“Bentires-Alj’s research team at the Department of Biomedicine, University of Basel and University Hospital Basel, elucidated one of these cascades. It begins with a metabolic enzyme called nicotinamide N-methyltransferase, or NNMT for short. And it ends with the substance that fills the space between the body’s cells and holds them together: collagen. Collagen is actually a good thing. But in the case of metastatic cancer, it betrays the body and helps cancer cells embed themselves in new tissues.”

““Triple negative” breast cancer, which affects roughly 15 percent of all breast cancer patients, is particularly aggressive because it often spreads throughout the body and forms lung and brain metastases. These breast cancer cells produce unusually high amounts of NNMT. [which] …causes the cancer cells to also produce more collagen than normal.” […]

“When the researchers removed NNMT from aggressive breast cancer cells and injected these cells into mice, the animals developed hardly any metastases. The cells also produced hardly any collagen.

A literature review also found that overproduction of NNMT is characteristic of a whole range of aggressive cancers – meaning it could be a universally important key factor in cancer metastasis.”

• Molecular Chain Reaction Helps Breast Cancer To Spread, June 7, 2023, Original story from the University of Basel, Technology Networks.

There is a history of research showing a role for NNMT in cancer survival or risk.

Abstract: “Nicotinamide N-methyltransferase (NNMT) has progressed from being considered merely a Phase II metabolic enzyme to one with a central role in cell function and energy metabolism. Over the last three decades, a significant body of evidence has accumulated which clearly demonstrates a central role for NNMT in cancer survival, metastasis, and drug resistance. In this review, we discuss the evidence supporting a role for NNMT in the progression of the cancer phenotype and how it achieves this by driving the activity of pro-oncogenic NAD+-consuming enzymes. We also describe how increased NNMT activity supports the Warburg effect and how it promotes oncogenic changes in gene expression. We discuss the regulation of NNMT activity in cancer cells by both post-translational modification of the enzyme and transcription factor binding to the NNMT gene, and describe for the first time three long non-coding RNAs which may play a role in the regulation of NNMT transcription. We complete the review by discussing the development of novel anti-cancer therapeutics which target NNMT and provide insight into how NNMT-based therapies may be best employed clinically.” (Parsons, Facey, 2021)

More background - overexpression of NNMT is also associated with obesity in addition to cancer:

Abstract: “Nicotinamide N-methyltransferase (NNMT) is a metabolic enzyme that catalyzes the methylation of nicotinamide (NAM) using the universal methyl donor S-adenosyl methionine (SAM), directly linking one-carbon metabolism with a cell's methylation balance and nicotinamide adenine dinucleotide (NAD⁺) levels. NNMT expression and activity are regulated in a tissue-specific-manner, and the protein can act either physiologically or pathologically depending on its distribution. While NNMT exerts a beneficial effect by regulating lipid parameters in the liver, its expression in adipose tissue correlates with obesity and insulin resistance. NNMT upregulation has been observed in a variety of cancers, and increased NNMT expression has been associated with tumor progression, metastasis and worse clinical outcomes. Accordingly, NNMT represents an appealing druggable target for metabolic disorders as well as oncological and other diseases in which the protein is improperly activated.” (Roberti, Fernández, Fraga, 2021)

Reduction of NNMT shows protective effects against cancer growth and spread:

Abstract excerpt: “Nicotinamide N-methyltransferase (NNMT) is a metabolic enzyme involved in controlling methylation potential, impacting DNA and histone epigenetic modification. NNMT overexpression has been described in various solid cancer tissues and even body fluids, including serum, urine, and saliva. Furthermore, accumulating evidence has shown that NNMT knockdown significantly decreases tumorigenesis and chemoresistance capacity.

»Most importantly, the natural NNMT inhibitor yuanhuadine can reverse epidermal growth factor receptor tyrosine kinase inhibitor resistance in lung cancer cells.” (Li, et al., 2022)

MicroRNA 449a seems to have regulatory control over NNMT expression.

The NNMT inhibitor yuanhuadine is mentioned by Bach, et al., 2018 also, identifying upregulation of microRNA 449a as a mechanism of action:

Abstract: “Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are used clinically as target therapies for lung cancer patients, but the occurrence of acquired drug resistance limits their efficacy. Nicotinamide N-methyltransferase (NNMT), a cancer-associated metabolic enzyme, is commonly overexpressed in various human tumors. Emerging evidence also suggests a crucial loss of function of microRNAs (miRNAs) in modulating tumor progression in response to standard therapies. However, their precise roles in regulating the development of drug-resistant tumorigenesis are still poorly understood.

Herein, we established EGFR-TKI-resistant non-small-cell lung cancer (NSCLC) models and observed a negative correlation between the expression levels of NNMT and miR-449a in tumor cells. Additionally, knockdown of NNMT suppressed p-Akt and tumorigenesis, while re-expression of miR-449a induced phosphatase and tensin homolog (PTEN), and inhibited tumor growth.

»Furthermore, yuanhuadine, an antitumor agent, significantly upregulated miR-449a levels while critically suppressing NNMT expression. These findings suggest a novel therapeutic approach for overcoming EGFR-TKI resistance to NSCLC treatment.” (Bach, et al., 2018) *paragraph breaks added by me.

What is yuanhuadine?

“yuanhuadine (1), a daphnane diterpenoid from the flowers of Daphne genkwa”. (Hong, et al., 2011)

Yuanhuadine is a diterpenoid which might be fragrant. That means it might activate G-protein coupled odor receptors and G-protein coupled receptors can have physiological functions outside of the nose like bitter taste receptors have roles beyond the tongue.

Daphne genkwa, the plant is mentioned to be poisonous but that the flower buds have been used medicinally. (landscapeplants.oregonstate.edu)

microRNA 449 may have effects by actions on SIRT1.

Pomegranate peel extract is consistently good at preventing cancer metastasis which is what led me to dig for microRNA connections to the NNMT findings in the Technology Network news story.

Modulation of microRNA can modulate SIRT1 (Wu, et al., 2022) *excerpt follows, and SIRT1 can modulate the activity of NNMT: “Sirt1 is required for the metabolic actions of Nnmt.” (Hong, S. et al., 2015) Which makes modulation of NNMT expression via SIRT1 modulation a possible mechanism of action for how pomegranate peel extract is so potent against cancer. Pomegranate peel extract has been shown to modulate SIRT4 via its actions promoting Nrf2:

Excerpt from my paper: Pomegranate promotes Nrf2, (Wang, T., et al, 2018) which promotes SIRT-4 which inhibits mast cell degranulation by inhibiting the metabolism of their mitochondria. (Hu B et al, 2020) Nrf2 and pomegranate peel inhibit mast cell degranulation. (Parisio, et al, 2020)

“(4) miRNAs play important roles in the regulation of SIRT1

miRNAs, a subtype of non-coding RNAs, are small endogenous RNAs which can inhibit protein translation in apoptosis.²²⁷ Moreover, SIRT1 has been revealed to be targeted by miRNAs such as miR-34a, miR-181, miR-128, miR-449 and miR-30a-5p. […] Other miRNAs, such as miR-449, have been investigated in a model of acute kidney injury model by detecting expression of its target SIRT1 and downstream factors p53/Bax.²³⁴ Inhibition of miR-449 elevated SIRT1 expression and inhibited acetylated p53 and Bax protein levels.^234” (Wu, et al., 2022)

And another section of my pomegranate paper about its benefits against cancer - consistently shown in many research studies:

• A literature review on pomegranate peel extract or juice and cancer treatment, primarily in vitro or in vivo studies. Whole fruit versus arils alone “has an enormous impact” on the polyphenol content and antioxidant potential. More clinical trials are needed. Liver and prostate cancer cells seem to be inhibited in growth with no negative effects on normal cells. Inhibits MAP Kinases, PI3K/AKT/mTor, and NF-κB, reducing:

1. inflammation related to CO-2 and iNOS,

2. cell proliferation related to PCNA and Ki67,

3. angiogenesis related to VEGF and CD-31, and

4. metastasis related to MMPs. (Figure 2, Syed, et al, 2013)

Identifying other NNMT inhibitors is also the focus of a review by Gao, et al., 2019:

“Other notable findings were the results obtained with the reference compounds. The general methyltransferase inhibitors, sinefungin and SAH, showed inhibitory activities in line with those previously reported. (24)” (Gao, et al., 2019)

Disclaimer: This information is being shared for educational purposes within the guidelines of Fair Use and is not intended to provide individual health guidance.

Reference List

• Molecular Chain Reaction Helps Breast Cancer To Spread, June 7, 2023, Original story from the University of Basel, Technology Networks.

(Bach, et al., 2018) Bach DH, Kim D, Bae SY, Kim WK, Hong JY, Lee HJ, et al., Targeting Nicotinamide N-Methyltransferase and miR-449a in EGFR-TKI-Resistant Non-Small-Cell Lung Cancer Cells. Mol Ther Nucleic Acids. 2018 Jun 1;11:455-467. doi: 10.1016/j.omtn.2018.03.011. Epub 2018 Mar 29. PMID: 29858080; PMCID: PMC5992482. https://www.cell.com/molecular-therapy-family/nucleic-acids/fulltext/S2162-2531(18)30042-8

(Gao, et al., 2019) Yongzhi Gao, Matthijs J. van Haren, Ed E. Moret, Johannes J. M. Rood, Davide Sartini, Alessia Salvucci, et al., Bisubstrate Inhibitors of Nicotinamide N-Methyltransferase (NNMT) with Enhanced Activity, J. Med. Chem. 2019, 62, 14, 6597–6614, July 2, 2019, https://doi.org/10.1021/acs.jmedchem.9b00413 https://pubs.acs.org/doi/10.1021/acs.jmedchem.9b00413

(Hong, et al., 2011) Hong JY, Chung HJ, Lee HJ, Park HJ, Lee SK. Growth inhibition of human lung cancer cells via down-regulation of epidermal growth factor receptor signaling by yuanhuadine, a daphnane diterpene from Daphne genkwa. J Nat Prod. 2011 Oct 28;74(10):2102-8. doi: 10.1021/np2003512. Epub 2011 Sep 14. PMID: 21916433. https://pubmed.ncbi.nlm.nih.gov/21916433/

(Hong, S. et al., 2015) Hong, S., Moreno-Navarrete, J., Wei, X. et al. Nicotinamide N-methyltransferase regulates hepatic nutrient metabolism through Sirt1 protein stabilization. Nat Med 21, 887–894 (2015). https://doi.org/10.1038/nm.3882 https://www.nature.com/articles/nm.3882

(Hu B et al, 2020) Hu B, Zhao S, Huang M, Ren J. ‘Nuclear factor E2 related factor (NRF2) inhibits mast cell- mediated allergic inflammation via SIRT4-mediated mitochondrial metabolism’. Ann Palliat Med. 2020 Nov;9(6):3839-3847. doi: 10.21037/apm-20-1848. Epub 2020 Nov 16. PMID: 33222465. Available at: https://pubmed.ncbi.nlm.nih.gov/33222465/

(Li, et al., 2022) Li XY, Pi YN, Chen Y, Zhu Q, Xia BR. Nicotinamide N-Methyltransferase: A Promising Biomarker and Target for Human Cancer Therapy. Front Oncol. 2022 Jun 9;12:894744. doi: 10.3389/fonc.2022.894744. PMID: 35756670; PMCID: PMC9218565. https://www.frontiersin.org/articles/10.3389/fonc.2022.894744/full

(Parisio, et al, 2020) Parisio, C., Lucarini, E., Micheli, L., et al., (2020) ‘Pomegranate Mesocarp against Colitis-Induced Visceral Pain in Rats: Effects of a Decoction and Its Fractions’. Int J of Molecular Sciences. 2020; 21(12):4304. https://doi.org/10.3390/ijms21124304 Available at: https://www.mdpi.com/1422-0067/21/12/4304/htm

(Parsons, Facey, 2021) Parsons RB, Facey PD. Nicotinamide N-Methyltransferase: An Emerging Protagonist in Cancer Macro(r)evolution. Biomolecules. 2021 Sep 28;11(10):1418. doi: 10.3390/biom11101418. PMID: 34680055; PMCID: PMC8533529. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8533529/

(Roberti, Fernández, Fraga, 2021) Roberti A, Fernández AF, Fraga MF. Nicotinamide N-methyltransferase: At the crossroads between cellular metabolism and epigenetic regulation. Mol Metab. 2021 Mar;45:101165. doi: 10.1016/j.molmet.2021.101165. Epub 2021 Jan 14. PMID: 33453420; PMCID: PMC7868988. https://www.sciencedirect.com/science/article/pii/S2212877821000053?via%3Dihub

(Syed, et al, 2013) Syed DN, Chamcheu JC, Adhami VM, Mukhtar H. Pomegranate extracts and cancer prevention: molecular and cellular activities. Anticancer Agents Med Chem. 2013;13(8):1149–1161. Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4052369/?tool=pmcentrez

(Wang, T., et al, 2018) Wang, T., Men, R., Hu, M., et al., (2018). ‘Protective effects of Punica granatum (pomegranate) peel extract on concanavalin A-induced autoimmune hepatitis in mice’. Biomed Pharmacother. Apr;100:213-220. doi: 10.1016/j.biopha.2017.12.110. Epub 2018 Feb 9. Available at: https://www.ncbi.nlm.nih.gov/pubmed/29428670

(Wu, et al., 2022) Wu, QJ., Zhang, TN., Chen, HH. et al. The sirtuin family in health and disease. Sig Transduct Target Ther 7, 402 (2022). https://doi.org/10.1038/s41392-022-01257-8, https://www.nature.com/articles/s41392-022-01257-8