Episomal-Persistent DNA in Cancer- and Chronic Diseases

Despite significant progress in cancer therapy, the lack of etiologic understanding in cancer causation and immunology, including synergistic (infectious) risk factors expected for >50% of cancers, limits current transfer from basic research to real-world application. As a continuation of the division of Harald zur Hausen, our group focuses on the interplay between Bovine Meat and Milk Factors (BMMF, more than 200 of such single-stranded, plasmid-like DNA elements are currently known) taken up by consumption of bovine nutrition products and the contribution to indirect induction of different types of cancer (pathologic model of ‘BMMF-induced zoonotic-linked, infectious, indirect carcinogenesis’) based on chronic inflammation (CI), induction of radicals and random DNA mutations. We also focus on BMMF in the context of chronic diseases of the human brain CNS. We apply both BMMF type-specific, ultrasensitive wet-lab as well as large-scale multi-omics in silico BMMF screening of publicly available DNA/RNA libraries to investigate the presence of BMMF as pathogenic risk factor in human cancer and in natural bio-reservoirs. In preclinical BMMF in vitro models, we test replication, transcription and expression of BMMF targets with the aim to generate and validate tools for BMMF detection and modulation of BMMF positivity.

More than 14 intensively-tested, DKFZ-built antibodies directed against a conserved BMMF protein (Rep) are used in streamlined, histochemical lab-based analysis pipelines in colorectal cancer (CRC), but also in lung, liver, pancreas and prostate cancer, amongst others, to assess diagnostic use and cancer-specific BMMF expression levels in CI-linked cancers and their (possible) inflammatory pre-cancer stages (e.g. diverticulitis, pancreatitis, non-alcoholic steatohepatitis (NASH), chronic obstructive pulmonary disease (COPD)). Causative contribution of BMMF to cancer, in particular to CRC, is assessed in histochemistry in a case vs. control and metric setup including pre-cancerous stages accessible via our broad clinical cooperation network. We combine it with additional protein-specific (immunoblotting and mass spectroscopy) and DNA/RNA-based readouts (qPCR/PCR, NGS, laser microdissection). In addition, we focus on the prognostic relevance of BMMF expression based on an immunoreactive BMMF score to evaluate BMMF expression in tumour and peritumour tissues and the associated outcome. We complement that with bulk as well as single-cell RNAseq large data analyses to test the effect of BMMF expression on the regulation of BMMF-positive macrophages to unravel the molecular and mechanistical background of BMMF-driven indirect carcinogenesis. This also includes the pro-tumourigenic modulation of the tumor microenvironment (TME) and therapy resistance, which might be supported by BMMF. We analyze the ultrastructure of BMMF targets (by immuno electron microscopy), which suggests a pleomorphic, vesicular presence of BMMF and accessibility for intervention - e.g. based on antibodies.

Our multi-omics read outs are steadily revised and expanded for new targets to unravel the role of BMMF in indirect carcinogenesis, which most likely represent one of the most common zoonotic-linked infections in humans. We expect a penetration of BMMF into 30-50% of current cancer cases and 40-60% of cancer deaths.

To foster cancer prevention based on BMMF technology, we analyze the effects of BMMF exposure in pre-clinical models (BMMF mouse model, cell (co-)culture models). Our vision is to establish early cancer diagnosis and systemic and/or BMMF antibody and immune cell therapy to reduce the burden of BMMF and to prevent/therapize BMMF-linked cancer.
 

• INTRODUCTION
• IN THE BEGINNING…
• CURRENT RESEARCH
• SIGNIFICANT ACCOMPLISHMENTS
• FUTURE OUTLOOK
• PHOTO GALLERY

Cecere, N., Alikhanyan, K., Ernst, C., Häfele, L., Feuerbach, L., Grewe, I., Siqin, S., Gunst, K., Shukla, G., Frehtman, V., Tessmer, C., Eichhorn, F., Allgäuer, M., Brobeil, A., Hofmann, I., Leuchs, B., Schneider, M.A., Bund, T. Bovine Meat and Milk Factor (BMMF) protein expression coincides with peritumor alveolar macrophages and is increased in lung cancer patients.
Submitted.

Siqin, S., Rahbari, M., Ernst, C., Grewe, I., Gunst, K., Neßling, M., Kaden, S., Richter, K., Häfele, L., Feuerbach, L., Krunic, D., Tessmer, C., Hofmann, I., Birgin, E., Brobeil, A., Rahbari, N., Heikenwälder, M., Bund, T. Expression of bovine meat and milk factor in hepatocellular carcinoma and colorectal liver metastasis patients. 
Submitted.

Siqin, S., Nikitina, E., Rahbari, M., Ernst, E., Krunic, D., Birgin, E., Tessmer, C., Hofmann, I., Rahbari, N., Bund, T. Bovine Meat and Milk Factor (BMMF) protein is expressed in macrophages spread far over the mucosa of colorectal cancer patients. 
Cells, in review.

Nikitina, E., Burk-Körner, A., Wiesenfarth, M., Alwers, E., Heide, D., Tessmer, C., Ernst, C., Krunic, D., Schrotz-King, P., Chang-Claude, J., von Winterfeld, M., Herpel, E., Brobeil, A., Brenner, H., Heikenwalder, M., Hoffmeister, M., Kopp-Schneider, A., Bund, T. Bovine meat and milk factor protein expression in tumor-free mucosa of colorectal cancer patients coincides with macrophages and might interfere with patient survival.
Molecular Oncology Volume 18, Issue 5, 2023. DOI: 10.1002/1878-0261.13390

Nikitina, E., Alikhanyan, K., Nessling, M., Richter, K., Kaden, S., Ernst, C., Seitz, S., Chuprikova, L., Haefele, L., Gunst, K., Rahbari, N., Birgin, E., Rasbach, E., Rahbari, M., Brobeil, A., Schenk, M., Büchler, M., de Villiers, E.-M., Bund, T.*, zur Hausen, H. Structural Expression of BMMF in tissues of colorectal, lung and pancreatic cancer patients.
International Journal of Cancer, Jul 1;153(1):173-182, 2022. (*corresponding author) DOI: 10.1002/ijc.34374 

Bund, T., Nikitina, E., Chakraborty, D., Ernst, C., Gunst, K., Boneva, B., Tessmer, T., Volk, N., Brobeil, A., Weber, A., Heikenwalder, M., zur Hausen, H., de Villiers, E.-M. Chronic Inflammatory Lesions of the Colon for BMMF Rep Antigen Expression and CD68 Macrophage Interactions.
Proceedings of the National Academy of Science U S A 118(12):e2025830118, 2021. DOI: 10.1073/pnas.2025830118

zur Hausen, H., de Villiers, E.-M. Bovine Meat and Milk Factors (BMMFs): Their Proposed Role in Common Human Cancers and Type 2 Diabetes Mellitus.
Cancers 13(21):5407, 2021. DOI: 10.3390/cancers13215407

de Villiers, E.-M., Gunst, K., Chakraborty, D., Ernst, C., Bund, T., zur Hausen, H. A specific class of infectious agents isolated from bovine serum and dairy products and peritumoral colon cancer tissue. 
Emerging microbes & infections 8:1205-1218, 2019. DOI: 10.1080/22221751.2019.1651620

Kilic, T., Popov, A.N., Burk-Körner, A., Koromyslova, A., zur Hausen, H., Bund, T.*, Hansman, G.S.* Structural analysis of a replication protein encoded by a plasmid isolated from a multiple sclerosis patient. 
Acta Crystallographica Section D, Structural biology 75:498-504, 2019. (*corresponding authors) DOI: 10.1107/S2059798319003991

zur Hausen, H., Bund, T., de Villiers, E.-M. Specific Nutritional Infections Early in Life as Risk Factors for Human Colon and Breast Cancers Several Decades Later. 
International Journal of Cancer, 144:1574-1583, 2019. DOI: 10.1002/ijc.31882

Eilebrecht, S., Hotz-Wagenblatt, A., Sarachaga, V., Burk, A., Falida, K., Chakraborty, D., Nikitina, E., Tessmer, C., Whitley, C., Sauerland, C., Gunst, K., Grewe, I., Bund, T. Expression and replication of virus-like circular DNA in human cells. 
Scientific Reports, 8:2851, 2018. DOI: 10.1038/s41598-018-21317-w

zur Hausen H., Bund T. and de Villiers, E.-M. Infectious Agents in Bovine Red Meat and Milk and Their Potential Role in Cancer and Other Chronic Diseases. 
Current Topics in Microbiology and Immunology, 407, 83-116, 2017. DOI: 10.1007/82_2017_3

Falida, K., Eilebrecht, S., Gunst, K., zur Hausen, H., de Villiers, E.-M. Isolation of Two Virus-Like Circular DNAs from Commercially Available Milk Samples.
Genome Announcements, 5(17):e00266-17, 2017. DOI: 10.1128/genomeA.00266-17.

zur Hausen, H. and de Villiers, E.-M. Dairy cattle serum and milk factors contributing to the risk of colon and breast cancers. 
International Journal of Cancer, 137(4), 959-67, 2015. DOI: 10.1002/ijc.29466

zur Hausen, H. What do breast and CRC cancers and MS have in common? 
Nature Reviews Clinical Oncology, 12(10), 569-70, 2015. DOI: 10.1038/nrclinonc.2015.154

zur Hausen, H. and de Villiers, E.-M. Cancer „causation“ by infections – individual contributions and synergistic networks. 
Seminars Oncology 41(6), 860-875, 2014. DOI: 10.1053/j.seminoncol.2014.10.003

Lamberto, I., Gunst, K., Müller, H., zur Hausen, H., and de Villiers, E.-M. Mycovirus-Like DNA Virus Sequences from Cattle Serum and Human Brain and Serum Samples from Multiple Sclerosis Patients. 
Genome Announcements, 2014, 2(4):e00848-14. DOI: 10.1128/genomeA.00848-14

Gunst, K., zur Hausen, H., and de Villiers, E.-M. Isolation of Bacterial Plasmid-Related Replication-Associated Circular DNA from a Serum Sample of a Multiple Sclerosis Patient.
Genome Announcements, 2014, 2(4):e00847-14. DOI: 10.1128/genomeA.00847-14

Whitley, C., Gunst, K., Müller, H., Funk, M., zur Hausen, H., and de Villiers, E.-M. Novel Replication-Competent Circular DNA Molecules from Healthy Cattle Serum and Milk and Multiple Sclerosis-Affected Human Brain Tissue.
Genome Announcements, 2(4):e00849-14, 2014. DOI: 10.1128/genomeA.00849-14. 

Funk, M., Gunst, K., Lucansky, V., Müller, H., zur Hausen, H., and de Villiers, E.-M. Isolation of Protein-Associated Circular DNA from Healthy Cattle Serum.
Genome Announcements, 2(4):e00846-14, 2014. DOI: 10.1128/genomeA.00846-14.

zur Hausen H., de Villiers, E.-M. Prenatal Infections with Subsequent Immune Tolerance Could Explain the Epidemiology of Common Childhood Cancers.
World Cancer Report, 2014.

Borkosky, S.S., Whitley, C., Kopp-Schneider, A., zur Hausen, H., de Villiers, E.-M. Epstein-Barr virus stimulates Torque-Teno virus replication: a possible relationship to multiple sclerosis. 
PLoS One, 7(2):e32160, 2012. DOI: 10.1371/journal.pone.0032160.

zur Hausen H. Red meat consumption and cancer: Reasons to suspect involvement of bovine infectious factors in colorectal cancer.
International Journal of Cancer, 130(11):2475-83, 2012. DOI: 10.1002/ijc.27413.