HUMAN PAPILLOMAVIRUS (HPV) AND PROSTATIC CANCER OSTEOBLASTIC BONE METASTASIS: HPV E6 CONTAINS AN ENDOTHELIN ACTIVE SITE TRYPTOPHAN

ARTP 18 Nov 2015 Paris Porte Maillot

TRAN Guy Mong Ky

Retired, Public Health (Agence Régionale de Santé ARS Auvergne Rhône Alpes), Hospital Hôtel-Dieu, Clermont-Ferrand, FRANCE. Correspondence: 31 Avenue du Bois 92290 Chatenay Malabry. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it . Phone: +33 9 81 89 38 70.

BACKKROUND Endothelin (ET-1, 2, 3) (Yanagisawa M, 1989) is mitogenic for 3T3 fibroblasts (Takuwa N, 1989), induces osteoblast proliferation and is involved in osteoblastic bone metastases of Prostatic Cancer (PK) with translocation TMPRSS2:ERG gene fusion (Delliaux C, 2014). We found earlier by centering on the HQLL motif that:

HPV-18  E2           IQTLNHQVL

contains the osteoprotegerin active site           ET  SHQLL

as well as parathormone (PTH)                           IQ LMHNL

and  PTH-related Protein (PTHrP)                        S EHQLL

that could explain PK osteoclastic bone metastasis (Tran GMK, 2004). We search for an endothelin in HPV that could explain osteoblastic bone metastasis.

METHODS Amino acid sequences and three dimensional structure (3D) comparison between HPV and ET.

RESULTS By centering on the crucial tryptophan W21, which removal decreases ET potency by a factor > 1000 (Kimura S, 1988), we found that

HPV (cand 89), -150, -160 E6                  34-EL-35         48-LDIVW-52  is homologous to the

ET-1,-2,-3/sarafotoxin active site                5-DM-6          17-LDI IW-21

In 3D structure, HPV E6 has 2 separated functional sites, centered on the COOH-terminal tryptophane W 21 and on 5-DM-6 (of sarafotoxine). The homologous sequence LNVVW is found in HPV-3, -28, -29, -68 ME 180. In HPV-18 E6, W was replaced by Y. Burrowing asp Attractaspis engaddensis sarafotoxin, which induces a coronary spasm, differs slightly from ET:QDVIW (where Q replaced L).

Weak but highly selective ET-A Receptor antagonists were isolated from Streptomyces Misakiensis and sp n° 7338 (Miyata S, 1992). Highly potent and selective ET-A Receptor antagonist structures are pentapeptides:

cyclo (D-Asp-Pro-D-Val-Leu-D-Trp) (Ihara M, 1992) and

cyclo (D-Asp-Pro-D-Ile -Leu-D-Trp) (D=Dextrogyre) (Lippton, 1993).

CONCLUSION The molecular homology of HPV E6 with the active site of Endothelin, implicated in PK bone metastasis, supports further the HPV role in KP, not only in the primary tumor, but also specifically in osteoblastic bone metastasis. Interestingly, high doses of Vitamin C has an anti-Endothelin action (Dow C, 2015).

BIBLIOGRAPHY Delliaux C, Endothelin-1, a gene regulated by TMPRSS2:ERG fusion proteins in prostate cancer bone metastases. ARTP, Paris, Nov 19 2014. Ihara M. Biological profiles of highly potent novel endothelin antagonists selective for the ETA receptor. Life Sci 1992, 50: 247-55. Dow C.Vitamin C Supplementation Reduces ET-1 System Activity in Overweight and Obese Adults.14th Int Conf on Endothelin: Physiology, Pathophysiology and Therapeutics. Press release of the Am Physiol Soc, 4th September 2015, www.the-aps.org/mm/hp/Audiences/Public-Press/2015/44.html. Kimura S. Structure-activity relationships of endothelin: importance of the C-terminal moiety. Bio Bio Res Com 1988, 156: 1182-6. Miyata S. WS-7338, new endothelin receptor antagonists isolated from Streptomyces sp. No. 7338. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological activities. J Antibiot (Tokyo) 1992, 45: 74-82; 83-7; 698-703;788-91; 1029-40; 1041-6. Lippton. Am Heart Assoc, 66^th Scientific Sessions, Georgia World Congress, Nov 8-11, 1993. Takuwa N. A novel vasoactive peptide endothelin stimulates mitogenesis through inositol lipid turnover in Swiss 3T3 fibroblasts. J Biol Chem 1989, 264: 7856-61. Tran GMK, Role of human papillomavirus type 18 in a subgroup of prostatic cancer with bone metastasis: Its protein E2 contains the osteoprotegerin active site EuroConf Cancer, Pasteur Inst. Jan 15-16 2004. Yanagisawa M. Molecular biology and biochemistry of the endothelins. Trends Pharmacol Sci 1989, 10: 374-8. Review.

PROSTATIC CANCER AND HUMAN PAPILLOMAVIRUS (HPV): MOLECULAR HOMOLOGY OF HPV-18 E2 WITH ONCOGENE CRIPTO-1.

Présenté à

ARTP 18 Nov 2015 Paris Porte Maillot

TRAN Guy Mong Ky

Retired, Public Health (Agence Régionale de Santé ARS Auvergne Rhône Alpes), Hospital Hôtel-Dieu, Clermont-Ferrand, FRANCE. Correspondence: 31 Avenue du Bois 92290 Chatenay Malabry. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it . Phone: +33 9 81 89 38 70.

BACKROUND HPV are found in Prostatic Cancer (PK) by PCR only when the primer is E (=Early) and not L (=Late) (Tran GMK, 2004). This particularity may be explained by L loss during integration (Johansson H, 2013). Cripto-1 (Teratocarcinoma-Derived Growth Factor-1), a member of Epidermal Growth Factor (EGF) family, is involved in PK progression (Terry S, 2013, 2015). It is a co-receptor of TGF-β, induces E-Cadherin loss (Zhong XY, 2008) in Epithelial to Mesenchymal Transition (Thiery JP, 2002) and stimulates the PI3K/Akt pathway. E-Cadherin down regulation can also be induced by HPV E6 (Matthews K, 2003). As HPV contains many oncogenes implicated specifically in PK [HPV-11 E2 = EGF-HRG-TGFα mitogens chimera (Tran MKG, 1997) with a common MHIESL hexapeptide, Osteoprotegerin in bone metastases (Tran GMK, 2004), Nucleophosmin 1 (NPM1) in androgeno-regulation (Tran GMK, 2014)], we search for a molecular homology between Cripto-1 and HPV.

METHODS Amino acid sequences comparison of HPV with Cripto-1 EGF motif        

                                                             CcxxGGxCxxxxxCxCxxxxxxxxC.

RESULTS

HPV-18 E2                                   113-CFkKGGqTV 45-YFdGNK D 127-DGNKDNC

Cripto-1                                         83-CL

..

A      S                                           D W T L      323-PPNN   L  KCWR  Q RC    122-VQVYF

matches with Cripto-1                        120-D(W,T)L      124-LPKKcsLcKCWHgQlRC    142-PQAFL

CONCLUSION The discovery of Cripto-1 being a viral HPV-18 E2 oncogene, whereas EGF is a HPV-11 E2, confirms the HPV role in PK, and E-cadherin loss as a factor of metastasis. A safe and efficient vaccine is warranted, as Gardasil and Gardasil 9 based on L1 may be followed by menopause/sterility and  multiple sclerosis, and Cervarix by thrombocytopenia. A BT PHARMA vaccine based on a HPV-16 and -18 oncogene E7, instead of L1 as in Gardasil and Cervarix, is in phase II trial at the Pasteur Institute (Paris). E7 is much smaller than L1 and has less worrying epitopes.

BIBLIOGRAPHY Matthews K. Depletion of Langerhans cells in human papillomavirus type 16-infected skin is associated with E6-mediated down regulation of E-cadherin. J Virol 2003, 77: 8378-85. Terry S. Involvement of CRIPTO-1 in progression of prostate tumors. Eurocancer, Paris, June 25-26, 2013. Terry S. CRIPTO overexpression promotes mesenchymal differentiation in prostate carcinoma cells through parallel regulation of AKT and FGFR activities. Oncotarget 2015, 6: 11994-2008. Thiery JP.  Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer 2002, 2: 442-54. Tran MKG. Human Papillomavirus (HPV) E2 protein contains a chimera of epidermal growth factor (EGF) and EGF family mitogens: Heregulin and Tumor Growth Factor. 6th Eur HIV Conf, Hamburg 1997, P474. Tran GMK. Role of human papillomavirus type 18 in a subgroup of prostatic cancer with bone metastasis: Its protein E2 contains the osteoprotegerin active site. EuroConf Cancer, Pasteur Inst. Jan 15-16 2004. Tran GMK. Prostatic cancer of viral origin: Homology of human oncogenic papillomavirus (HPV) L1 with nucleophosmin (NPM1), a controller of androgen receptor transcription. ARTP, Nov 19, 2014, Paris. Zhong XY. Positive association of up-regulated Cripto-1 and down-regulated E-cadherin with tumour progression and poor prognosis in gastric cancer. Histopathology. 2008, 52: 560-8.

PROSTATIC CANCER AND HUMAN PAPILLOMAVIRUS (HPV): AUTO-IMMUNE THROMBOCYTOPENIA INDUCED BY GARDASIL AND CERVARIX VACCINE HPV L1 MIMICKING PLATELET GPV

Présentation 

ARTP 18 Nov 2015 Paris Porte Maillot

TRAN Guy Mong Ky

Retired, Public Health (Agence Régionale de Santé ARS Auvergne Rhône Alpes), Hospital Hôtel-Dieu, Clermont-Ferrand, FRANCE. Correspondence: 31 Avenue du Bois 92290 Chatenay Malabry. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it . Phone: +33 9 81 89 38 70.

BACKROUND Among the auto-immune complications occurring after anti-HPV vaccination (Gardasil and Cervarix), was reported thrombocytopenia (Pugnet G, 2009). Odds Ratio (OR) for Serious Autoimmune Adverse Events (SAAEs) reported by the Vaccine Adverse Event Reporting System (VAERS) from Jan 2006 through Dec 2012 is OR=1,3 (95% CI=0.48-3.5) (Geier DA, 2015) for thrombocytopenia. If severe, this thrombocytopenia can conduct to cerebral hemorrhage and eventually death.

METHODS Amino acid sequences comparison between HPV L1 and platelet gpV. We centered the alignment on the FP and related LP motifs (Leucine L replaces Proline P). Platelet alloantigen, P1T, on glycoprotein V is associated with neonatal alloimmune thrombocytopenia (Ertem M, 1994). Measles was investigated versus platelet gpIIbIIIa on the FP motif.

RESULTS   Platelet gpV was aligned with HPV (types -16,-18, -6, -11,-31,-33,-52,-58) L1 of Gardasil 9.

perfect molecular mimicry ALPDG was found between HPV-61,-72,-81 L1 and platelet gpV.

A common motif  LPDT was found between rubella virus nsp and HPV-18 L1. Rubella can induce thrombocytopenia (Okazaki N, 2011). There is a common motif ALPD between gpV, HPV-11 and HPV-6  L1. Anti-gpV specific antibodies are also particularly evident in Varicella virus thrombopenia (Mayer JL, 1996) and are cross-reactive with platelet (Wright JF,1994): Varicella ORF 24 and rubella nsp share the ALPDT motif.

There was a perfect molecular mimicry between Measles P protein strain Mvi/Victoria.AUS/12.99 (Bankamp B, 2008) and platelet gpIIb: 

   platelet gpIIb                                                                         782-RGNSFP-787

Measles P protein strain Mvi/Victoria.AUS/12.99                      199-RGNSFP-204

The other Measles virus strains have                                              RGNNFP

CONCLUSION  The perfect 100% molecular mimicry ALPDG, centered on a Proline, between HPV-61,-72,-81 L1 and platelet gpV raises the question of the etiology of the so-called "idiopathic" thrombocytopenic purpura. It is advocated to research HPV-61,-72,-81 as possible culprits, either by PCR or Metagenomics (Johansson H, 2013). HLA-DR3 may be a  risk factor. The alimentation must be strictly controlled, avoiding in particular aspirin and alliaceous. The obligatory vaccination by Measles Mumps Rubella vaccine, associated with Zoster-Varicella vaccine (Okazaki N, 2011) and 3 doses of Gardasil or Gardasil 9 or Cervarix, may enhance further the auto-immune response against platelet gpV and induce autoimmune thrombocytopenia. Gardasil 9 is particularly worrying.

BIBLIOGRAPHY Bankamp B. Genetic variability and mRNA editing frequencies of the phosphoprotein genes of wild-type measles viruses. Virus Res 2008, 135: 298-306. Ertem M. A new platelet alloantigen, P1T, on glycoprotein V associated with neonatal alloimmune thrombocytopenia. Pediatr Res 1994, 35: 160A. Geier DA. A case-control study of quadrivalent human papillomavirus vaccine-associated autoimmune adverse events. Clin Rheumatol 2015, 34: 1225-31. Johansson H. Metagenomic sequencing of "HPV-negative" condylomas detects novel putative HPV types. Virology 2013, 440: 1-7. Mayer JL. Varicella-associated thrombocytopenia: autoantibodies against platelet surface glycoprotein V. Pediatr Res 1996, 40: 615-9. Meenaghan M. Antibodies to platelet glycoprotein V in polytransfused patients with haematologic diseases. Vox Sang 1993, 64:167–70. Okazaki N. Detection of platelet-binding anti-measles and anti-rubella virus IgG antibodies in infants with vaccine-induced thrombocytopenic purpura. Vaccine 2011, 29: 4878–80. Pugnet G. Immune thrombocytopenic purpura following human papillomavirus vaccination. Vaccine 2009, 27: 3690. Wright JF. Virus-reactive antibodies cross-react with autologous platelets in a patient with varicella zoster virus (VZV)-associated idiopathic thrombocytopenic purpura (ITP). Blood 1994, 84 (suppl):185a

GARDASIL ANTI-PAPILLOMAVIRUS VACCINE: MOLECULAR MIMICRY OF HPV L1 WITH GONADOLIBERIN GNRH EXPLAINING PRECOCIOUS MENOPAUSE AND STERILITY

TRAN Guy Mong Ky

Retired, Public Health (Agence Régionale de Santé Auvergne Rhône Alpes), Hospital Hôtel Dieu, Clermont-Ferrand FRANCE. Correspondence: 31 Avenue du Bois 92290 Chatenay Malabry. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it . Phone: +33 9 81 89 38 70.

Association pour la Recherche sur les Tumeurs de la Prostate (ARTP)

18 Nov 2015 . Palais des Congrès, Porte Maillot, Paris