Understanding the cell cycle - chemotherapy and beyond Julie Mycroft Principal Pharmacist Paediatric Oncology Royal Marsden NHS Trust Cancer treatment modalities Can be used alone or in combination Outcome measured in terms of survival rates and response rates Aims of treatment Chemotherapy To reduce tumour burden Improvement in symptoms Eradication of metastases → cure Hormone Therapy Manipulate hormone environment leading to regression of tumours sensitive to hormones Adjuvant setting Palliation in advanced disease Neoadjuvant History of chemotherapy development 1946 Nitrogen mustard given to treat lymphomas 1947 Antifolates introduced 1949 Methotrexate introduced 1950s 5-Fluoro-uracil synthesised 1952 6-mercaptopurine described 1954 Actinomycin D introduced 1960s Combination chemo cured childhood ALL and HD Recent Years Many new agents Focus changes to optimising timing and usage and modulating toxicity The cell cycle Phases of the cell cycle G0 resting phase G1 early growth phase S DNA synthesis G2 later growth phase M Mitosis Cell division – mitosis (1) Prophase Chromatin condenses into chromosomes. Each chromosome duplicates and consists of 2 sister chromatids. Nucleus breaks down Metaphase Chromosomes align and are held by microtubules attached the mitotic spindle and to the centromere Cell division – mitosis (2) Anaphase The centromeres divide. Sister chromatids separate and move toward the corresponding poles Telophase Daughter chromosomes arrive at the poles and the microtubules disappear. The condensed chromatin expands and the nuclear envelope reappears The cytoplasm divides, the cell membrane pinches inwards and two daughter cells are produced The Cell Cycle and Tissue Growth The rate of cell division in human tumours varies considerably from one disease to another Majority of common cancers increase in size slowly compared to sensitive normal tissues such as BM and GI epithelium The relationship between cell cycle and cell death affects tumour growth Chemotherapy Effects Cytotoxic drugs produce their effects by damaging the reproductive potential of cells The more rapidly growing tumours are more likely to respond to drug treatment this accounts for leukaemias, lymphomas and testicular cancers being more responsive than colonic / pancreatic cancers Growth Fraction At a given time, the number of cells in a population that are actively passing through the cell cycle divided by the total number of cells in the population = growth fraction The greater the growth faction, the more likely the treatment will produce cell death Kinetics of cell killing Fractional Cell kill hypothesis A given dose of cytotoxic drug kills a given proportion of cells, not a given number Smaller tumours require fewer cycles of chemotherapy than larger ones Pulsed intermittent therapy Maximises tumour cell killing whilst allowing normal tissues damaged by the drug to recover Cytotoxic Drug Classification Cell Cycle Phase-Specific Agents active in a particular phase of cell cycle Depend on the production of some type of unique biochemical blockade of a particular reaction occurring in a single phase of the cell cycle Cell Cycle Phase-Non-specific Agents Cytotoxic effect exerted irrespective of cell cycle state equally effective in large tumours in which cell growth is low dose dependent single dose has same effect as repeated fractions totalling the same amount Phase Specificity of Cytotoxic Drugs Phase of cell cyle Effective agents G1 Steroids, asparaginase S phase Antimetabolites G2 Bleomycin, etoposide Mitosis Vinca alkaloids, taxanes Phase non-specific Alkylating agents, nitrosoureas, antibiotics, procarbazine, dacarbazine, platinums Mechanisms of Action (1) Alkylating agents and nitrosureas Highly reactive molecules Interfere with replication by covalently linking an alkyl group (R-CH2+) to nucleic acids and proteins of the base pairs of the cellular DNA causes the strands of DNA to cross-link either within a strand or between strands. Mechanism of toxicity - impairment of DNA replication Examples e.g cyclophosphamide, chlorambucil, melphalan, nitrosoureas eg carmustine alkylator-like agents - cisplatin, carboplatin, procarbazine, dacarbazine Mechanisms of Action (2) Antitumour Antibiotics Disrupt normal replication by binding to DNA intercalating between the base pairs blocking the transcription of DNA Breaks in DNA may also occur Examples Anthracyclines ie Doxorubicin Epirubin, Mitoxantrone, Actinomycin D Bleomycin, Mechanisms of Action (3) Antimetabolites Interfere with normal synthesis of nucleic acids Cell cycle phase specific (S phase) folate antagonist - MTX pyrimidine antagonists cytarabine, 5-fluorouracil, capecitabine, gemcitabine purine antagonists cladribine, mercaptopurine, thioguanine, fludarabine adenosine deaminase inhibitor - pentostatin Mechanisms of Action (4) Disrupt the M phase of the cell cycle Vinca alkaloids Inhibit the assembly of microtubules by binding to tubulin resulting in the dissolution of the mitotic spindle required for chromosome division e.g vincristine, vinblastine, vinorelbine Taxanes Bind to stabilised microtubules once they have formed, resulting in arrest of normal mitotic cell division and subsequently cell death e.g paclitaxel, docetaxel Mechanisms of Action (5) Camptothecans Inhibit type I DNA topoisomerase. Act predominantly in the S phase e.g topotecan, irinotecan Epipodophyllotoxins Inhibit type II DNA topoisomerase and prevent cells from entering mitosis Produce proteinassociated DNA double strand breaks e.g etoposide Mechanisms of Action (6) – Misc. Asparaginase Normal Cell Tumor Cell Aspartic Acid + L-Glutamine L-Asparagine Synthetase L-Asparagine (cell produced) L-A´ase L-Asparagine Deficiency Aspartic Acid + Ammonia Cell Proliferation Cell Death Combination Chemotherapy (1) Early studies used single agents, but remissions were short and relapse was associated with drug resistance Combination chemotherapy is used to try and improve rate and duration of response by combining drugs with different mechanisms of action. This also helps prevent resistance mechanisms Despite knowledge of cell kinetics, most regimens have been decided on empirically Combination Chemotherapy (2) The combination of drugs is chosen based on some common principles Use drugs that are known to be effective as single agents Use drugs with non-overlapping toxicity pulsed intermittent therapy should be used to allow the GIT and the bone marrow to recover Use the optimal dose and schedule for each individual agent If possible use drugs with synergistic killing effects Use drugs which work at different phases in the cell cycle Follow schedules that are supported by experience or observation, not just theory Short-term toxicity Haematopoietic System Gastro-intestinal tract Bone marrow suppression occurs when the pool of stem cells has been damaged by chemotherapy. The store of mature blood cells in the bone marrow lasts for around 8 to 10 days following treatment, after which leucopenia and thrombocytopenia can develop Nausea and vomiting are common in patients treated with intravenous alkylating agents, doxorubicin and cisplatin. Hair loss Scalp cooling may be used Long term toxicity Impaired gonadal function Pulmonary fibrosis Busulfan, Bleomycin Organ damage Procarbazine and alkylating agents men – decreased spermatogenesis women – ovarian failure Liver damage – antimetabolites Cardiac damage – anthracyclines Second Cancers Alkylating agents, etoposide, anthracyclines Drug Resistance Cells in a solid tumour are not uniformly sensitive to a cytotoxic drug As the tumour grows, greater heterogeneity develops and cell mutation occurs Host defence mechanisms and the use of cytotoxic drugs exert a selection pressure encouraging the survival of the resistant cells , which grow and multiply Cellular Mechanisms of Resistance Mechanism Drug Efficient repair to damaged DNA alkylating agents Decreased uptake by cell MTX, doxorubicin Increased drug efflux (p-glycoprotein) epipodophyllotoxin vincs, anthracyclines Decreased intracellular activation 6MP,5-FU Increased intracellular breakdown cytarabine Bypass biochemical pathways MTX, 6MP, asparaginase Gene amplification or overproduction of blocked enzyme MTX, nitrosoureas Other Mechanisms of Drug Resistance Diminished vascularity Only a small proportion of cells may be in cycle, allowing time for repair from cytotoxic damage before cell division Hormone Therapy Beatson demonstrated that some inoperable breast cancers regressed after removal of the ovaries (oophorectomy) Many years later Huggins showed that metastatic prostatic cancer regressed after removal of the testes (orchidectomy). In breast cancer, hormone receptor status is clinically important in management Principals of hormone therapy (1) Receptor proteins for steroid hormones are found in both the cytoplasm and the nucleus The interaction between these hormones and their receptor proteins promotes cell growth and division. The steroid hormone crosses the cell membrane and forms a complex with a receptor in the cytoplasm. This activated complex passes into the nucleus where it binds to a protein, leading to the production of messenger RNA (mRNA) and then protein. Finally DNA is synthesised and the cell divides Principals of hormone therapy (2) This interaction provides the rationale for a number of ways in which hormone manipulation can modify tumour growth. It may be possible to: lower the plasma concentration of hormone by removing the source of production, e.g. the testes or ovaries prevent the hormone from binding to receptor via competitive inhibition or by reducing synthesis of the receptors block binding of the hormone/receptor complex to DNA in the nucleus The precise mode of action of agents used in hormone therapy is often unclear. Approaches to hormone therapy (1) Lowering plasma hormone concentration Radiotherapy Surgery Radiotherapy to the ovaries induces the menopause – the ovaries stop producing eggs and the female sex hormones. In breast cancer can involve: the ovaries (oophorectomy) the adrenals (adrenalectomy) the breast tissue (mastectomy) In prostate cancer: surgical removal of the testes (orchidectomy) Medical treatment Aromatase inhibitors Analogues of luteinizing hormone–releasing hormone (e.g. goserelin and leuprorelin) Approaches to hormone therapy (2) Blocking the action of circulating hormones Anti-oestrogens and anti-androgens work by blocking the binding of hormones to their receptors. Anti-oestrogens (e.g. tamoxifen) Anti-androgens (e.g. flutamide, megestrol acetate) Additive hormone therapies The action of circulating hormones can also be blocked by additive hormone therapies, which in breast cancer include oestrogens, androgens, glucocorticoids and progestogens. Summary A knowledge of the cell cycle is important to understanding the mechanism of action of cancer chemotherapy Combination chemotherapy is used to try and improve rate and duration of response by combining drugs with different mechanisms of action. Manipulating the interaction between hormones and cell growth provides a means for treating hormone sensitive cancers Acknowledgements The Institute of Cancer Research - interactive education unit module 4 “An approach to therapies” ‘Cancer and it’s Management’ - 3rd Edition Souhami R and Tobias J (Blackwell Science)