Salauddin Al Azad and Md. Iqbal Khan. HAMLET (Human Alpha- Copyright© Salauddin Al Azad and Md

Advances in Pharmacology and Clinical Trials ISSN: 2474-9214

HAMLET (Human Alpha-Lactalbumin Made Lethal to Tumor Cells) - A Hope for the Cancer Patients

Salauddin Al Azad* and Md. Iqbal Khan Mini Review Biotechnology and Genetic Engineering Discipline, Life Science School, Khulna Volume 4 Issue 1 University, Bangladesh Received Date: March 01, 2019 Published Date: March 18, 2019 *Corresponding author: Salauddin Al Azad, Biotechnology and Genetic Engineering DOI: 10.23880/apct-16000152 Discipline, Life Science School, Khulna University, Khulna, Bangladesh, Email: [email protected]

Abstract

Cancer is a malignant disease which is in most part incurable. It is not curable through normal medication. Different cure techniques have already equipped to wipe cancer out but still a mess that cancer finds its own way-out. Oncolytic viral therapy is a new promising strategy against cancer. Oncolytic viruses (OVs) can replicate in cancer cells but not in normal cells, leading to lysis of the tumor mass but are recognized by the immune system as pathogens and the consequent anti- viral response could represent a big hurdle for OVs makes the concept compromised for cancer treatment or malignant metastasis. Every human cell has a hereditary program that upon enactment will cause cell demise, named apoptosis. Cancer cells can develop because of imbalanced expansion, cell cycle guideline and their apoptosis hardware: genomic mutant particles bringing about non-practical professional apoptotic proteins or over-articulation of against apoptotic sister proteins which structure the premise of tumor development. Shockingly, exercises gained from infections demonstrate that malignancy can't be viewed essentially as the inverse of apoptosis. Using anticancer genes as a therapy for cancer can be effective as they can go through the gene lines and make a call for destruction of malignancy. HAMLET (Human Alpha-lactalbumin Made Lethal to Tumor Cells) is such an anticancer gene which is found in human milk that can be effective in cancer treatment and also refusing new path making of cancer.

Keywords: HAMLET; Oncolytic Viruses; Apoptosis; P53; HDI; MAPK

Abbreviations: Hamlet: Human Αlpha-Lactalbumin Introduction Made Deadly to Tumor Cells; MAL: Multimeric Alpha- Lactalbumin; OV: Oncolytic Viruses; Bcl2: B-Cell Anticancer genes are proto-oncogenes when become Lymphoma 2; HDI: Histone Deacetylase Inhibitor; oncogenic ectopically overexpressing, explicitly annihilate P21waf1: Cyclin-Dependent Kinase Inhibitor 1; MAPK: tumor cells without hurting typical cells [1]. This phone Mitogen Activated Protein Kinase; Gtpase: Guanosin Tri obliteration emerges for an assortment of systems, for Phospatease; P53: Phosphoprotein 53. example, apoptosis, mitotic disaster pursued by apoptosis or corruption, and autophagy.

HAMLET (Human Alpha-Lactalbumin Made Lethal to Tumor Cells) - A Hope for the Cancer Patients Adv Pharmacol Clin Trials

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Anticancer qualities rose up out of concentrates on xenograft demonstration in patients with skin papillomas disease cells in the late 1990s. Viruses were found over a and in patients with bladder malignancy. The instruments century back however from early occasional ailments like of tumor-cell-passing stay hazy, be that as it may. disease and particularly leukemia was endeavored to be treated with infections. All through the written history of Following the experience with tumor cells, HAMLET illnesses, there have been perceptions of malignant attacks the cells what's more, causes mitochondrial layer growth relapse upon characteristic co-disease with depolarization, cytochrome c discharge, infections [2]. HAMLET is a conceivable chemotherapeutic phosphatidylserine presentation, and a low caspase operator with the capacity to kill malignant growth cells. reaction. A small amount of the cells experiences Alpha-lactalbumin is the essential protein part of human morphological changes normal for apoptosis, yet caspase milk. In a recent report, it has revealed that multimeric restraint does not protect the cells and Bcl-2 alpha-lactalbumin (MAL), a compound confined from a overexpression or modified p53 status does not impact on small amount of human milk called casein, incited what the affectability of tumor cells to HAMLET. HAMLET gave off an impression of being apoptosis in human lung additionally makes a condition of unfurled protein over- carcinoma cells, pneumococcus and different pathogens, burden and enacts 20S proteasomes, which adds to cell while leaving solid, separated cells [2]. It has been the demise. In parallel, HAMLET translocates to tumor cell ideal fix in this case. The dynamic segment in charge of cores, where high-fondness collaborations with histones the tumoricidal movement was observed to be a complex cause chromatin interruption, loss of translation, and of alpha-lactalbumin and oleic acid. Its activity is the atomic buildup. The diminishing cells moreover show opposite of the oncogens. HAMLET (Human Alpha- morphological changes perfect with macro-autophagy lactalbumin Made Lethal to Tumor Cells) is a sub-atomic and later ponders demonstrate that macro-autophagy is complex got from human milk that kills tumor cells by a associated with the cell passing reaction to HAMLET. The procedure taking after apoptosis. The intricate comprises outcomes propose that HAMLET which is similar to a of mostly unfurled Alpha-lactalbumin and oleic corrosive, hydra with numerous heads, may connect with a few and both the protein with the unsaturated fat are significant cell organelles, in this way actuating a few required for cell demise. HAMLET has expansive types of cell demise, in parallel. This multifaced nature antitumor activity in vitro, and its restorative impact has may underlie the quick passing reaction of tumor cells been affirmed in vivo in a human glioblastoma rodent and the expansive antitumor action of HAMLET.

Figure 1: HAMLET is defined as the product of partially unfolded Alpha-lactalbumin and oleic acid.

Through programmed cell death the human body trigger tumor cell demise. HDIs trigger tumor cell demise dispenses with undesirable cells without bringing out an by reestablishing the outflow of antitumor qualities, inflammatory reaction [3,4]. Cells experiencing though HAMLET ties histones with high fondness and customized cell death notifying an unmistakable changes the chromatin structure. This ponder thought morphological appearance described by cell shrinkage, about the impacts of HAMLET on tumor cell feasibility, membrane blistering, atomic buildup, and fragmentation histone acetylation, and DNA trustworthiness when the with the arrangement of ‘apoptotic bodies’; this sort of antagonists were joined. HDI pretreatment caused a cell passing is for the most part referred to as apoptosis stamped increment in the demise reaction to HAMLET [3,4]. HAMLET alters the structure of chromatin and and HAMLET a further increment in the histone

Salauddin Al Azad and Md. Iqbal Khan. HAMLET (Human Alpha- Copyright© Salauddin Al Azad and Md. Iqbal Khan. Lactalbumin Made Lethal to Tumor Cells) - A Hope for the Cancer Patients. Adv Pharmacol Clin Trials 2019, 4(1): 000152.

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hyperacetylation reaction to HDIs. Cell demise was joined to trigger cell passing by means of the extraneous, demise by expanded DNA harm and diminished p21WAF1 receptor– subordinate pathway and by means of the articulation when the two agonists were consolidated. We inborn mitochondrial demise pathway through actuation recommend that the cooperative energy depends on or hindrance of explicit Bcl-2 relatives, by means of the various however uniting demise pathways, which include direction of responsive oxygen species and through cell modifications of the chromatin. As HDIs and HAMLET cycle [5]. HAMLET acts straightforwardly on the have archived remedial impacts, the joined utilization of mitochondria furthermore, triggers an effector caspase HAMLET and HDIs may be of an incentive in the reaction and the atomic caspase 2 reaction to HAMLET treatment of malignant growth. Tumor cell passing may add to assist mitochondrial enactment. The demise of expanded particularly when HDIs and HAMLET were HAMLET-treated cells is caspase independent, in any case joined and the impact was synergistic when TSA was [6,7]. In the present examination, the DNA harm reaction included prior to HAMLET. The sub-atomic premise of was appeared to be caspase-free. supporting the idea that this cooperative energy is misty, yet a few passing traditional apoptosis isn't the way to the joined impact of pathways might be talked about. Caspases are basic for HAMLET and TSA. the execution of apoptosis and HDIs have been proposed

Reaction mode Activity References The potentiality of HAMLET based treatment of human A. Apoptosis mimic mode of action provoked glioblastoma cells in brain xenograft through mechanisms [8] by HAMLET mimic to apoptosis have proved clinically in recent years Investigation for proteins with blocking properties of adhesion of bacteria to cancer cells have already commenced B. Simultaneous anticancer and antibacterial and instead one of the milk proteins exposed with actual [9] activity of HAMLET killing power of cancer cells while leaving the normal cells unharmed The apoptotic response that accompanies death in HAMLET C. HAMLET has been shown to target skin treated tumor cells and the role of mitochondria in this papillomas and bladder cancers process have analyzed through progressive inhibition of MAPK kinases and GTPases D. HAMLET act as molecular signal modulator to the APC mutated cells in cancer HAMLET proved itself as hyperactive antagonist to colon accumulating in tumour tissue precisely and cancer with APC mulation in individuals as a new, peroral

reducing β-catenin and related tumour agent markers Alpha-lactalbumin is a substrate specifier in lactose synthase E. Tumoricidal HAMLET complex can identify complex, showing its antitumor activity after partial conserved death pathways suitable for unfolding and binding to oleic acid molecule. This [10] targeting configuration unifies HAMLET as therapeutic molecular complex HAMLET is a complex of Alpha-lactalbumin and oleic acid F. Plasma membrane disruption of protein- which is also termed as liprotide, have positive starring in [11] lipid complex cancer cell membrane disposal Biosynthesis of nanoparticles (non-toxic to normal cells and tissues) act as a substitute of Alpha-lactalbumin in the G. HAMLET-like active molecule [12] liprotide (protein-lipid complex) where oleic acid plays the therapeutic role

H. HAMLET can be prepared from non-specific Programmed cell death and cancer specific cytotoxicity have sources but renders same effects in observed when Alpha-lactalbumin-oleic acid complex source [13,14] therapeutic questions is from bovine

Table 1: Analysis of different modes of activity of HAMLET against cancer cells.

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Like apoptin, HAMLET meddles with chromatin tumor-specific apoptosis in a p53-free way. Over- structures proposing that particularly these cell articulation of the counter apoptotic Bcl-2 and caspase structures are critical for its tumor-particular apoptosis restraint does not save HAMLET positive cells. As of late, acceptance [15]. HAMLET activatestes 20S proteasomes, have given proof that HAMLET initiates macro-autophagy which additionally adds to cell demise. HAMLET initiates [16, 17].

Clinical study on Clinical outcome [16]

8/9 patients showed tumor regression No adverse Bladder cancer reactions were reported 29/5 patients with complete cure No side effect Skin papilloma detected – p53-independent – Cytochrome c release – Activation of caspase pathways – Circumvents Bcl-2 over- Tumor-selective cell death expression & caspase inhibition – Interference with characteristics [16] chromatin structure – Located in nucleus, mitochondria, ER, & proteasomes – Induction of macro-autophagy

Table 2: Main Therapeutic and mechanistic cell death aspects of HAMLET.

Macro-autophagy is a lysosomal catabolic pathway 5. Bolden JE, Peart MJ, Johnstone RW (2006) Anticancer reusing cell particles. In malignancy cells, the macro- activities of histone deacetylase inhibitors. Nature autophagy pathways have been changed in contrast with reviews Drug discovery 5(9): 769-784. their ordinary partners (Høyer-Hansen and Jäättelä, 2008), which may clarify (to some degree) the watched 6. Hakansson A, Zhivotovsky B, Orrenius S, Sabharwal H, tumor-specific action of HAMLET. Future ponders must Svanborg C (1995) Apoptosis induced by a human be completed to recognize in detail the exact components milk protein. Proceedings of the National Academy of empowering HAMLET's tumor-particular apoptosis for all Sciences 92(17): 8064-8068. these tumor cells. 7. Hallgren O, Gustafsson L, Irjala H, Selivanova G, References Orrenius S, et al. (2006) HAMLET triggers apoptosis but tumor cell death is independent of caspases, Bcl-2 1. Hallgren O, Aits S, Brest P, Gustafsson L, Mossberg AK, and p53. Apoptosis 11(2): 221-233. et al. (2008) Apoptosis and tumor cell death in 8. Fischer W, Gustafsson L, Mossberg AK, Gronli J, Mork response to HAMLET (human Alpha-lactalbumin S, et al. (2004) Human Alpha-lactalbumin made lethal made lethal to tumor cells). Adv Exp Med Biol 606: to tumor cells (HAMLET) kills human glioblastoma 217-240. cells in brain xenografts by an apoptosis-like 2. Ho CSJ, Rydstrom A, Trulsson M, Baalfors J, Storm P, mechanism and prolongs survival. Cancer research et al. (2012) HAMLET: functional properties and 64(6): 2105-2112. therapeutic potential. Future oncology 8(10): 1301- 9. Marks LR, Clementi EA, Hakansson AP (2012) The 1313. human milk protein-lipid complex HAMLET sensitizes 3. Schwartz LM (1995) Changes in the Structure and bacterial pathogens to traditional antimicrobial Function of the Multicatalytic Proteinase agents. PloS one 7(8): e43514. (Proteasome) during Programmed Cell Death in the 10. Rath EM, Duff AP, Hakansson AP, Vacher CS, Liu GJ, et lntersegmental. al. (2015) Structure and potential cellular targets of 4. Wyllie AH (1987) Apoptosis: cell death in tissue HAMLET-like anti-cancer compounds made from milk regulation. The Journal of pathology 153(4): 313-316. components. Journal of Pharmacy & Pharmaceutical Sciences 18(4): 773-824.

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11. Frislev HS, Boye TL, Nylandsted J, Otzen D (2017) cancer therapy: Mechanisms of action. Molecular Liprotides kill cancer cells by disrupting the plasma Biology 49(2): 231-243. membrane. Scientific reports 7(1): 15129. 15. Backendorf C, Visser AE, De Boer AG, Zimmerman R, 12. Delgado Y, Morales-Cruz M, Hernández-Román J, Visser M, et al. (2008) Apoptin: therapeutic potential Hernández G, Griebenow K (2015) Development of of an early sensor of carcinogenic transformation. HAMLET-like cytochrome c-oleic acid nanoparticles Annu Rev Pharmacol Toxicol 48: 143-469. for cancer therapy. Journal of Nanomedicine & Nanotechnology 6(4): 1. 16. Aits S, Gustafsson L, Hallgren O, Brest P, Gustafsson M, et al. (2009) HAMLET (human α‐lactalbumin made 13. Zhong S, Liu S, Chen S, Liu H, Zhou S, et al. (2015) lethal to tumor cells) triggers autophagic tumor cell Cytotoxicity and apoptosis induction of bovine alpha- death. International journal of cancer 124(5): 1008- lactalbumin-oleic acid complex in human breast 1019. cancer cells. Food Science and Technology Research 21(1): 103-110. 17. Scarlatti F, Granata R, Meijer A, Codogno P (2009) Does autophagy have a license to kill mammalian 14. Lezhnin YN, Kravchenko YE, Frolova EI, Chumakov cells? Cell Death Differ 16(1): 12-20. PM, Chumakov SP (2015) Oncotoxic proteins in