Chemotherapy and Glycosylation Therapeutic Impact

Carcinogens are widely spread in developed countries that widened the different cancer types` epidemiology. Cancer treatment consists of four main categories, the surgical treatment in operable and early diagnosed cases, chemotherapy, immunotherapy and radiotherapy[1]. Chemotherapeutic agents such as alkylating agents, anti-metabolites, anti- tumor antibiotics , topoisomerase inhibitors, mitotic inhibitors and plant alkaloids are used for cancer cell cycle arrest, alteration metabolism and apoptosis. Generally, usage of chemotherapy is designed for neoadjuvant therapy before surgery to diminish its size and permits its operability. Also, chemotherapy has a palliative role in advanced cancer stages to improve life and reduce pain [2]. In my new treatment strategy, there will be specific chemotherapeutic drugs according to their liability to glycosylation and DNA damage. The most common group is glycosylated platinum.(IV) prodrugs such as oxaliplatin, cisplatin and oxaliplatin [3]. Most of chemotherapeutic agents can be carried by glycosylated nanoparticles as targeting molecules for avoidance severe side effects[4]. According to the previous role, many chemotherapeutic agents can be selected for that strategy. But why glycosylation is chosen? The answer of that question is included in the bad need of glycoside and glucose to malignant cells, as they are the fuel of the high rate anabolic demand of cancer cells through aerobic glycolysis by Warburg effect [5].Besides that, cancer cells are rich in glycoside transporters, membrane glycosides [6]. Also, glycosylated chemotherapeutic agent can be linked to carcinogenic proteins and transcriptional factors.

Role of Cancer Transcriptional Factors in Chemo-Biological Therapy

 Cancer transcriptional factors constitute a marvelous deceiving tool for cancer cells for uptake the biological or the chemotherapeutic agent and in the same time highly selective targeting for cancer cells only. There are many restrictions in usage such transcriptional factors and their structure must be changed in a smart and a complicated way. In other words, transcriptional factors should be common in many cancer types and upregulate many carcinogenic genes such as wnt5a and Hif-1α [7,8]. Also, many carcinogenic proteins that are commonly involved in cancer signaling cascades such as growth factor receptor tyrosine kinases (RTKs, EGFR), small GTPases (e.g., Ras), serine/threonine kinases (e.g., Raf and Akt), cytoplasmic tyrosine kinases (e.g., Src and Abl), lipid kinases (e.g., phosphoinositide 3-kinases, PI3Ks), as well as nuclear receptors (e.g., the estrogen receptor, ER) [9] are appropriate also. The reason for choosing that type of proteins because they are mutated forms then secreted from cancer cells. Their carcinogenic function is evident as they are a part of cancer signaling pathways within the tumor microenvironment. One of their most important advantages is favorism and great availability within the cancer extracellular matrix [10]. But if they are used by their same structures, they will spread malignancy even in healthy host cells.

Transcriptional Factors Post Translational Modifications in Chemo-Biological Therapy

So changing their structure is critical, however it is complicated. The suggested post-translational modifications of carcinogenic transcriptional factors are classified into three categories. The first is preservation the post translational modifications that keep the 3^rd structure of the protein as the same. The second is disabling the effector and the carcinogenic post-translational modification of the transcriptional factor and replacing it with a therapeutic molecule if possible. That step is critical because we have to link the therapeutic molecule to that modified amino acid by the same carcinogenic chemical group. The third is aiming to make the modified transcriptional factor have more affinity that the carcinogenic transcriptional factor ( author). For example, wnt5a is involved in many cancer signaling mechanisms (canonical, non canonical and Ca ⁺² dependent) [11, 12] and also associated with cellular senescence[13], tumor initiation from different cancer stem cells [14], chemotherapy resistance [15], β catenin upregulation in tumor microenvironment in pyllodes tumors [16], tumor-associated inflammation [17], epithelial mesenchymal transition [18],cell migration and invasion in nasopharyngeal carcinoma [19] and gastric carcinoma [20]. So, wnt5a is a common and familiar molecule to different cancer cells and components. When it is combined with cisplatin, the most important suggested post- translational modification is depalmitoylation Cysteine 104 because palmitoylation of cysteine 104 is included in many carcinogenic canonical and non-canonical pathways after its secretion [21]. In addition, GPI anchoring of serine 282 changed the wnt5a into an arrested signaling molecule [22]. The final step is glycosylation asparagine residues 114, 120, 311 and Asn325 with sialylated glycosides gave wnt5a molecule two advantages, the first is the better affinity than the carcinogenic wnt5a towards malignant cells, endothelial cells, leucocytes and platelets after binding the modified wnt5a to different selectins that are expressed on previously mentioned cells [23] . The second advantage is attachment 4 glycosylated cisplatin molecules on the four previously mentioned glycosylated asparagine residues. So, the modified wnt5a played a pivotal role in advancing the efficacy of cisplatin [3]. As a result, the modified wnt5a molecule after the previously mentioned post-translational modifications acts a dormant widespread carrier of 4 cisplatin molecules towards the primary tumor and the metastatic sites. In the end, it can be used in advanced cancer cases with the least side effects. [24]Another example is usage the modified Hif1α as a broad spectrum anti-cancer treatment, especially in solid tumors. Hif1α is related to many cancer genes upregulation (Hypoxia response element) [25] such as carbonic anhydrase IX, glucose transporter-1 (GLUT-1), that is a transmembranous glucose transporter [26]. GLUT-3, hexokinase 2, lactate dehydrogenase [27] and vascular endothelial growth factor (VEGF)in solid tumors [25], so it is a good candidate for our strategy.Nitrosylated cysteine 800 of Hif1α is crucial for hif-1α carcinogenic action [28]. So, the modified hif-1α will be denitrosylated biochemically by Flavohemoglobin within the culture medium [29], after then it will be blocked by attachment to carbonic anhydrase IX inhibitor (a therapeutic molecule for reversal the hypoxic tumor microenvironment, primary sulfonamide with NO moiety (2-methoxy-4-(nitroxy)butoxy)-3-oxoprop-1-en-1-yl)benzyl4-sulfamoylbenzoate), which acts as nitric oxide donor [30]) by the same original post- translational modification (nitrosylation) not to change the 3rd structure of hif1α Presence of nitric oxide donor is essential for S-nitrosylation by S100A9 [31] Serine residues 247, 551, 555, 576, 589 and 657 will be O-glycosylated with 6 glycosylated cisplatin molecules after phosphorylation inhibition. The previously mentioned phosphorylated serine residues will stabilize the hif-1α molecule which is not desired for the suggested function. Other post translational modifications are done to the modified hif-1α to be easily degradable such as desumoylation of lysine residues 391, 477, 532, 538, 547 [32]and deacetylation of lysine 709 [33]. When the modified hif-1α is degraded within the cancer cell, it will release carbonic anhydrase IX inhibitor and 6 glycosylated cisplatin molecules inside one malignant cell [34]. Here in, the importance of the modified transcriptional factor hif1αcomes by delivering concentrated dosage of chemotherapy to one cancer cell in a highly specific manner, especially in highly expressed hif1α cancer types such as breast, colon, gastric, lung, skin, ovarian, pancreatic, prostate, and renal cancer [35].

Monoclonal Antibodies Glycosylated Fc Region Binding To Modified Transcriptional Factors

 Immunotherapy has emerged the cancer therapy. It stimulates the human immune response against cancer by a memory treatment by B and T lymphocytes. Monoclonal antibodies against specific cancer antigens such ashuman epidermal growth factor receptor (HER)-1 and HER-2] or oncogenes (RAS] and VEGF evoked a reasonable efficacy with less side effects. In addition, it increased the phagocytic function of CAR-T cell [36]. Apparently, MABs seem a part of human immunity, but they are also blind, not selective as expected. Many MABs cause autoimmune self-response because the carcinogenic targeted antigens can be in healthy tissues also such as immune checkpoint inhibitors. They can cause inflammatory dermatitis, hepatitis , colitis, dermatomyositis and encephalitis [37] . MABs also showed poor responses and high relapse rate after long term treatment due to many complicated resistance mechanisms. For example, poor tumor immunogenicity and loss of neoantigen expression [39,40]. Also, genetic heterogeneity and enormous neoantigen expression overcome MABs efficacy [41]. Another resistance mechanism is cancer cell mutation of β2-microglobulin and down-regulation of MHC- class 1 , afterthen resistance immune checkpoint blockade [42]. So, we can conclude that malignant cell DNA heterogeneity is the most relevant resistant element.In chemo-biological therapy, we can select the main transcriptional factor that is responsible for resistance, then induce specific post-transcriptional modifications that preserve its structure, ban its carcinogenic effect by disruption the effector side group of the domain (amino acid) and prepare other side chains chemically to bind to chemotherapeutic molecule and finally attaches it to monoclonal antibody Fc region by a glycoside bond. For example, the immune check point inhibitor resistant molecule, which is STAT1. Phosphorylated STAT1 by IFN-γ activation of JANs kinases [43] or by IL-12 activation[44], and then enhance interferon- stimulated genes replications (ISGs), in the end, increase PD-1 transcription to overcome the number of immune check point inhibitors [45]. In the chemo-biological therapy, stat1 post-translational modifications will be designed as the following: phosphorylated tyrosine stat1 [46], with deaminated lysine residues [47]will be purified in HeLa cell after activation by IFN-γ, then nuclear inactivation occurs by unknown tyrosine phosphatase enzyme [48] to stop its transcriptional activity in HeLacells to yield stat1-PTP ( unphosphorylated stat1), afterthen E6-E6AP complex will degrade PTP from stat1-PTP complex in vitro to produce the final form (in vitro phosphorylated stat1) [49]. The Purified phosphorylated stat1 contains 7 deaminated lysine residues which are glycosylated by 7 A5 cisplatin ( glycosylated cisplatin molecules). Then, the glycosylated with be attached to solamargine Fb region of nivolumab (previously designed) by rhamnosyl transferase. The resulted compound will be phosphorylated stat1 which is translocated to the malignant cell nucleus containing 7 glycosylated cisplatin molecules, in turn will damage the malignant PD-1 genetic locus [50].

In the chemo-biological strategy of cancer therapy is divided into two components, the first one (the biological one) is cancer related modified protein with specific post-translational modifications that preserve its main 3rd structure, disable its carcinogenic effect and preparation of amino acids side chains for merging with a chemotherapy. The second element is the chemotherapeutic agent, but in concentrated form and highly selected towards the cancer cell nucleus, specifically to the responsible gene of the chemotherapeutic drug resistance after loading it on a familiar transcriptional factor (the first component). I think, we have to be more intelligent in fighting the cancer. That strategy will enable us to create many destructive malignant cell damaging drugs by the same cancer weapons.

There is no conflict of interest

That mini-review article is dedicated to the all suffering cancer patients all over the world with my wishes for complete cure.

1. DL. Cellular Mechanisms of Chemotherapy.2015.
2. Simon C, Watson M, Drake A, Fenton A, McLoughlin C. Principles of cancer treatment. Innov Ait. 2008; 1:107-118.
3. Ma J, Wang Q, Yang X, Hao W, Huang Z, Zhang J, et al. Glycosylated platinum(IV) prodrugs demonstrated significant therapeutic efficacy in cancer cells and minimized side-effects, 2016;45:11830-11838
4. Glycosylated Nanoparticles for Cancer-Targeted Drug Delivery.Sergio Andres Torres-Perez, Cindy Estefani Torres-Pérez, Sonia Mayra Perez-Tapia and Eva Ramon-Gallegos. Front. Oncol., 30 November 2020;10: - 2020.
5. The Warburg Effect: How Does it Benefit Cancer Cells?Maria V. Liberti1,2 and Jason W. Locasale2, Trends Biochem Sci. 2016; 41: 211–218.
6. Cai L, Gu Z, Zhong J, Wen D, Chen G, He L, et al. Advances in glycosylation-mediated cancer-targeted drug delivery. Drug Discovery Today. 2018; 23:1126-1138.
7. Wnt5a Signaling in Normal and Cancer Stem CellsYan Zhou, 1 Thomas J. Kipps, and Suping Zhang Stem Cells Int. 2017; 2017.
8. Hypoxia-Inducible Factors and Cancer. Jonathan C. Jun, AmanRathore, HarisYounas, Daniele Gilkes, and Vsevolod Y. Polotsky, Curr Sleep Med Rep. 2017; 3: 1-10.
9. Signal Transduction in CancerRichard Sever and Joan S. Brugge,Cold Spring Harb.Perspect Med. 2015; 5: [10] Cancer and the tumor microenvironment: a review of an essential relationship
10. Flaubert Mbeunkuiand Donald J. Johann, rCancer Chemother Pharmacol. 2009 Mar; 63: 571-582. Published online 2008; 14.
11. Amerongen RV, Nusse R. Towards an integrated view of Wnt signaling in development. Development. 2009; 136: 3205-3214.
12. Mikels AJ, Nusse R, Purified Wnt5a protein activates or inhibits β-catenin-TCF signaling depending on receptor context. PLoS Biol. 2006; 4:79.
13. Campisi J, di Fagagna FD. Cellular senescence: When bad things happen to good cells. Nat. Rev. Mol. Cell Biol. 2007; 8:729-740.
14. Akbari-Birgani S, Paranjothy T, Zuse A, Janikowski T, Cieslar-Pobuda A, Likus W, Urasinska E, Schweizer F, Ghavami S, Klonisch T. et al. Cancer stem cells, cancer-initiating cells and methods for their detection. Drug Discov. Today. 2016; 21:836-842.
15. Ramos P, Bentires-Alj M. Mechanism-based cancer therapy: Resistance to therapy, therapy for resistance. Oncogene. 2015; 34:3617-3626.
16. Sawyer EJ, Hanby AM, Rowan AJ, Gillett CE, Thomas RE, Poulsom R, Lakhani SR, Ellis IO, Ellis P, Tomlinson IP. The Wnt pathway, epithelial-stromal interactions, and malignant progression in phyllodes tumors. J. Pathol. 2002; 196:437-444.
17. Pereira C, Schaer DJ, Bachli EB, Kurrer MO, Schoedon G. Wnt5a/CaMKII signaling contributes to the inflammatory response of macrophages and is a target for the antiinflammatory action of activated protein C and interleukin-10. Arterioscler. Thromb. Vasc. Biol. 2008; 28:504510.
18. Jordan NV, Prat A, Abell AN, Zawistowski JS, Sciaky N, Karginova OA, Zhou B, Golitz BT, Perou CM, Johnson G.L. SWI/SNF chromatin-remodeling factor Smarcd3/Baf60c controls epithelial-mesenchymal transition by inducing Wnt5a signaling. Mol. Cell. Biol. 2013; 33:3011-025.
19. Qin L, Yin YT, Zheng FJ, Peng LX, Yang CF, Bao YN, Liang YY, Li XJ, Xiang YQ, Sun R, et al. Wnt5a promotes stemness characteristics in nasopharyngeal carcinoma cells leading to metastasis and tumorigenesis. Oncotarget. 2015; 6:10239-10252.
20. Kurayoshi M, Oue N, Yamamoto H, Kishida M, Inoue A, Asahara T, Yasui W, Kikuchi A. Expression of Wnt-5a is correlated with aggressiveness of gastric cancer by stimulating cell migration and invasion. Cancer Res. 2006; 66:10439-10448.
21. Schulte G, Bryja V, Rawal N, Castelo-Branco G, Sousa KM, Arenas E. Purified Wnt-5a increases differentiation of midbrain dopaminergic cells and dishevelled phosphorylation. J Neurochem. 2005;92: 1550-1553.
22. Wnt Signaling Is Regulated by Endoplasmic Reticulum RetentionJ. Susie Zoltewicz , Amir M. Ashique ,YoungshikChoe ,GenaLee,Stacy Taylor, KhanhkyPhamluong,MarkSolloway,Andrew S. Peterson. 2009
23. https://www.sciencedirect.com/science/article/pii/S0005273603003183
24. Atta WO. And Azeiz AZA. Wnt5a Role in Cisplatin Effect and Selectivity Augmentation in Cancer Therapy. Open Access Library J. 2021; 8: 1-12.
25. Tian H, McKnight SL, Russell DW. Endothelial PAS Domain Protein 1 (EPAS1), a Transcription Factor Selectively Expressed in Endothelial Cells. Genes & Development.1997; 11; 72-82.
26. Arany Z, Huang LE, Eckner R, Bhattacharya S, Jiang C, Goldberg MA, et al. An Essential Role for P300/CBP in The cellular Response to hypoxia. Proceedings of the National Academy of Sciences of the United States of America.1996; 93:12969-12973.
27. Arany Z, Huang LE, Eckner R, Bhattacharya S, Jiang C, Goldberg MA, et al. Bunn. 1996; 93:12969-12973.
28. Akt-Mediated Activation of HIF-1 in Pulmonary Vascular Endothelial Cells by S-Nitrosoglutathione.D. Jeannean Carver, Benjamin Gaston, Kimberly deRonde, and Lisa A. Palmer. Is J Respir Cell Mol Biol. 2007; 37: 255- 263.
29. Alexander DF, Judith F, Christian JT, Bollinger PTK. Bacterial Hemoglobins and falvohemoglobins for Alleviation of Nitrosative Stress in Escherichia coli. Applied and Environmental Microbiology. 2002; 68:4835-4840.
30. Carradori S, Mollica A, Monte CD, Ganese A. Supuran, CT. Nitric Oxide Donors and Selective Carbonic Anhydrase Inhibitors: A Dual Pharmacological Approach for the Treatment of Glaucoma, Cancer and Osteoporosis. Molecule. 2015; 20: 5667-5679.
31. Nancy RG. Knowing Where to S-Nitrosylate. Science Signaling, 2014; 7: 314.
32. Morris F, Gerald W. The Introduction of Enzymes into the Cells by Means of Liposomes. Journal of Lipid Research, 1978; 19: 289-303.
33. Seo KS, Park JH, Heo JY, Jing K, Han J, Min KN, Kim C, Koh GY, Lim K, Kang GY, Lee JU, Yim YH, Shong M, Kwak TH, kweon GR. SIRT2 Regulates Tumour Hypoxia Response by Promoting HIF-1α Hydroxylation. Oncogene.2014; 34:1354-1362.
34. Atta W, Azeiz A. Effect of Modified HIF-1α Linked to Carbonic Anhydrase IX Inhibitor and Glycosylated Cisplatin on Solid Tumors: Short Review. Journal of Biosciences and Medicines. 2021; 9, 59-72.
35. Dales JP, Garcia S, Meunier-Carpentier S, Andrac-Meyer L, Haddad O, Lavaut MN, et al. Overexpression of Hypoxia-Inducible Factor HIF-1α Predicts Early Relapse in Breast Cancer: Retrospective Study in a Series of 745 Patients. Int. J. 2005; 116: 734-739.
36. Schirrmacher V. Quo Vadis Cancer Therapy? Fascinating discoveries of the last 60 years. Lambert Academic Publishing; 201; 1-353.
37. Management of the Adverse Effects of Immune Checkpoint InhibitorsManuel Morgado, Ana Placido, Sandra Morgado, and FatimaRoqueVaccines . 2020;8: 575.
38. Mechanisms of Cancer Resistance to ImmunotherapyRilanBai 1, Naifei Chen 1, Lingyu Li 1, Nawen Du 1, Ling Bai 1, ZhengLv 1, HuiminTian 1, Jiuwei Cui 1 Front Oncol. 2020; 10:1290.
39. Schumacher TN, Schreiber RD. Neoantigens in cancer immunotherapy. Science. 2015; 348: 69-74.
40. Takeda K, Nakayama M, Hayakawa Y, et al. IFN-γ is required for cytotoxic T cell-dependent cancer genome immunoediting. Nat Commun. 2017; 8:14607.
41. Strickland KC, Howitt BE, Shukla SA. Association and prognostic significance of BRCA1/2-mutation status with neoantigen load, number of tumor-infiltrating lymphocytes and expression of PD-1/PD-L1 in high grade serous ovarian cancer. Oncotarget. 2016; 7: 13587-13598.
42. Zaretsky JM, Garcia-Diaz A, Shin DS. Mutations associated with acquired resistance to PD-1 blockade in melanoma. N Engl J Med. 2016; 375: 819-829.
43. Walter MR. The Role of Structure in the Biology of Interferon Signaling.Front. Immunol. 2020; 11: 606489.
44. IL-12 Family Cytokines in Cancer and ImmunotherapyBhalchandraMirlekar and YuliyaPylayeva-Gupta. Cancers (Basel). 2021;13:167.
45. Wang IM, Lin H, Goldman SJ, Kobayashi M. STAT1 is activated by IL-4 and IL-13 in multiple cell types. MolImmunol. 2004; 41: 873-84.
46. Kim TK, Maniatis T. Regulation of of interferon-γactivated STAT1 by the ubiquitin-proteosome pathway. Science. 1996; 273:1717-1779.
47. Munagala R, Aqil F, Jeyabalan J, Kandimalla R, Wallen M, Tyagi N, Wilcher S, Yan J, Schultz DJ, Spencer W, Gupta RC. exosome-mediated delivery of RNA and DNA for gene therapy. Cancer Lett. 2021; 505:58-72.
48. Hoeve JT, de Jesus Ibarra-Sanchez M, Fu Y, Zhu W, Tremblay M, David M, Shuai K. identification of a nuclear Stat1 protein tyrosine phosphatase. Mol Cell Biol. 2002; 22: 5662-5668.
49. Jing M, Bohl J, Brimer N, Kinter M, Vande Pol SB. Degradation of tyrosine phosphatase PTPN3 by association with oncogenic human papillomavirus E6 proteins. J Virol. 2007; 81: 2231-2239.
50. Atta WO. Usage of Nivolumab - Platinum Containing STAT1 Molecule for Suppression PD-1/PD-L1 Genes in PD-1/PD-L1 Expressing Cancer Cells. Global Journal of Medical Research. 2022; 22: 55-63.