MUNI SCI Bi4025en Molecular Biology Mgr. Jiří Kohoutek, Ph.D. 1 Department of Experimental Biology Lecture 10 • Molecular basis of carcinogenesis - oncogene versus suppressors 2 Department of Experimental Biology Tumor associated facts • Every third person in the Czech Republic gets cancer. • One in four people dies because of it. • In 2015, there were almost 542,000 people living in the Czech Republic who had been diagnosed with an oncological disease in that year or earlier. • Every year, about 27,000 people die of cancer in the Czech Republic. • Mortality from malignant tumors shows stagnation in absolute numbers! • Increase in prevalence (number of patients with a given tumor living in a given year). 3 Department of Experimental Biology MUNI SCI Incidence of malignancies in Czech Republic • Every 20 minutes, one person dies of cancer in the Czech Republic. • In the number of oncological patients - leading places in Europe. • The most common newly diagnosed malignant diseases in 2011-2015 were skin tumors (excluding melanoma), colorectal cancer, breast cancer in women, and malignant tumors of the prostate and lung. • Main factors responsible for high incidence of malignant disease in Czech Republic: o Significant demographic aging of the Czech population, o New treatment technologies and their success: prolonging survival of cancer patients, o Subsequent malignancies in oncological patients. Duseketal 2018 MUNI 4 Department of Experimental Biology _ _ _ H H yy NOR 2017 g Q Incidence of malignancies as a global problem • Tumors 12% of all deaths (56 million in 2000). • In 2000 - 5.3 million men and 4.7 million women were diagnosed, 6.2 million died of tumor. • Prediction - 10 million new cases in 2000 to 15 million in 2020 - aging population - smoking and poor lifestyles. 5 Department of Experimental Biology Update: http://www.who.int/topics/cancer/en/ MUNI SCI Incidence of malignancies as a global problem Estimated number of new cases in 2020, worldwide, both sexes, all ages Other cancers 8 879 843 (46%) Cervix uteri Liver 604 127 (3.1%) 905 677 (4.7%) Total : 19 292 789 Data source: Globocan 2020 Graph production: Global Cancer Observatory (hr.trj://gco.iarcir) l"!e;r:aVonal Ane-;y RW«»rch Wl ClIKif 6 Department of Experimental Biology MUNI SCI Important facts About 1/3 of tumors are associated with obesity, poor diet and low physical activity. Smoking - 30% of all deaths from tumors, 87% of deaths from lung tumors. Obesity increases the risk of breast tumors in postmenopausal women by 50% and 40% in bowel tumors in men. 30% Smoking and alcohol use (172,000 deaths due to smoking and 19,000 deaths due to alcohol use.) 18-20% Chronic infections^ {Deaths occur mostly in poor countries due to hepatitis B virus, human papillomavirus, HIV, human T-cell leukemia^ lymphoma, and,, others.) 18-20% Hormones 30-35% Unbalanced diet {One-third of all cancer deaths due to too many high-glycemic carbohydrates, too many calories leading to obesity, and lack of physical activity.) •1% „ * Pollution Occupation (DeatnS0CCur mostly where pollution is heavy.) 7 Department of Experimental Biology MUNI SCI _ Cancers and rates Global cancer rates The ten worst countries and the UK cancer rate, per 100,000 of the population 1 Denmark 326.1 2 Ireland 317 3 Australia 314.1 4 New Zealand 309.21 5 Belgium 306.81 6 France 300.4 1 7 US 300.21 8 Norway 299.11 9 Canada 296.6 | 10 Czech Republic 295 § 22 UK 266.9 ^| SOURCE- WORLD CANCER RESEARCH FUND 8 Department of Experimental Biology HIGHEST CANCER RATES IN THE WORLD Coses per 100. OQOptopie CANADA 297 rRELAND 317 NORWAY AUSTRALIA ZEALAND 299 314 309 BELGIUM DENMARK 307 326 Tumor • Uncontrolled growth of cells in the tissue of higher organisms. • Does not have a physiological function. • Clonal expansion characteristics. • Disrupts the balance in the body. • Critical two types of genes: • Oncogenes (gain of function). • Tumor suppressors (loss of function). oAll of these genes generally have other primary functions. oGenes may be tumor suppressor or oncogene in one tissue, at one time point, NOT in other. oEffects of such genes are subject to tradeoffs with other functions. MUNI 9 Department of Experimental Biology r> r» t Tumor oncogenes • 1970 - src- chicken retrovirus Rous sarcoma virus (Dr. Martin, UC Berkley). • 1976 - Stehelin, Vermus, Bishop - oncogenes are activated proto-oncogenes. • Proto-oncogene - a gene that encodes proteins that affect growth, differentiation and/or, signal transmission. • Protooncogene activation - is the conversion of proto-oncogene into oncogene. Mutation or increased expression or amplification - oncogene Ras, Myc, ERK. • Proto-oncogene mutations are: • Activating. • Dominant. • Occur in somatic cells and rarely in germ cells. MUNI 10 Department of Experimental Biology r> r» t Tumor oncogenes ANTI MITOGEN IC SIGNALS MITOGÉNJC SIGNALS ONCOGENESIS E E Id2 promotor Normal development MUNI 11 Department of Experimental Biology https://blogs.shu.edu/cancer/2014/12/23/merck-acquires-oncoethix-for-novel-myc-blocker/ O U J. Tumor suppressors • The products of genes for tumor suppressors (antioncogenes) in normal cells do not cause proliferation, but, on the contrary, suppress it and keep the cells at rest (GO). Their loss is manifested by unregulated proliferation. • Mutations of tumor suppressors are: • Inactivating^ • Recessive (associated with Loss Of Heterogeneity) ("recessive oncogenes"). • Occur in somatic and also in germ cells. 12 Department of Experimental Biology MUNI SCI Tumor suppressor p53 - guardian of the genome. • 17p13.1, 53 kDA. • Proliferation, apoptosis, DNA repair, angiogenesis, replication, cell division... • Rapid degradation. • Regulated MDM2 protein - transfer to cytoplasm, inactive. • Induction of expression after cell exposure to stress. • Cell cycle arrest, apoptosis. • Mutations in more than 50% of human tumors. • Li Fraumeni syndrome - hereditary tumor syndrome, germ mutation p53. • Mutations of inactivating, recessive, somatic and germ cells. 13 Define footer - presentation title / department MUNI SCI p53 Signaling Extracellular space Tumor suppressor Hypoxia / I Ionizing Radiation Chemotherapy TNF. Fast. TRAIL I ROCKLAND flLZL antibodies & assays 14 Department of Experimental Biology MUNI SCI Tumor suppressor miR-192 miR-194 miR-215 miR-221 miR-605 miR-17-3p miR-25 miR-32 miR-143 miR-145 miR-18b miR-193a miR-339-5p miR-509-5p miR-660 miR-661 mi R-191 mi R- 10a miR-34a miR-661 miR-887 let-7 miR-885-3p MDMX miR-125b miR-1285 miR-141 miR-380-5p miR-504 miR-92 miR-15a miR-16 miR-25 miR-30d miR-453 miR-98 miR-200a miR-19b miR-518c miR-638 MicroRNAs in the regulation of p53 and mutants n_ ■— mTOR I— TSC1 1- miR-32 pathway 1 RPL11 4— Ribosomal stress 15 Department of Experimental Biology https://www.frontiersin.org/articles/10.3389/fonc.2015.00284/full MUNI SCI Types of genes mutated in cancers • Gatekeeper genes - genes that regulate growth and differentiation; include oncogenes and tumor suppressor genes. • Caretaker genes - genes that help to maintain genetic integrity; their loss of function mutations lead to: o Microsatellite instability (due to mismatch repair deficiency). o Chromosomal instability (gain or loss of chromosomes or parts thereof). • (3) Landscaper genes - genes that when mutated lead to abnormal extracellular or intracellular environment that contributes to carcinogenesis. 16 Department of Experimental Biology MUNI SCI Risk factors • The risk of developing a tumor increases with increasing age. • Exposure to chemicals. • Viruses. • Mutations. • Lifestyle? • Early diagnosis and treatment important. • Identification of persons at risk. 17 Department of Experimental Biology Journal of Nucleic Acids, January 2010, (6551):592980 MUNI SCI Chemicals increase risk of cancer • Asbestos. • Pesticides. • Herbicides. • Benzene. • Radioactive substances. • Chemicals in food - herbicides, pesticides, E-numbers. 18 Department of Experimental Biology MUNI SCI Oncogenic (Cancerogenic) viruses • Retroviruses (RNA viruses): contain an oncogene (acutely transforming viruses) in their genome or activate the proto-oncogene next to which they have integrated (slowly transforming). • DNA oncogenic viruses use a different transformation strategy: they do not contain oncogenes, but encode proteins that interact with tumor suppressors (RB, p53, p300/CBP) of the host cell and thus push the host cell into the S phase: • SV40 - large T antigen interacts with p53, RB, p300/CBP through different domains. • Adenovirus - E1A interacts s RB a p300/CBP; E1B associates s p53. • Papilomavirusus HPV-16, HPV-18 - E6 interacts with p53, p300/CBP; E7 interacts with RB. 19 Department of Experimental Biology MUNI SCI Oncogenic viruses and human diseases • RNA viruses: • human lymphotropic virus type I (HTLV-1) causes adult T-cell leukemia (ATTL) • DNA viruses: • Epstein-Barr Viruses (EBV) - Burkitťs lymphom (BL), Hodgkine lymphom (HD), Lymphoms, Nasofaryngial carcinomes (NPC). • Hepatitis B virus (HBV) - Hepatocelular carcinom (HCC). • Human papillomaviruses (HPV 16, 18,..) - anogenital tumors, oral tumors, warts. • Human herpesvirus type 8 (HHV8) - Kaposi sarcoma (KS). 20 Department of Experimental Biology MUNI SCI Historical perspective • 400 BC Hippocrates described tumors as long prominences. • Grece: karkinos = crayfish; onkos = crab • Latin cancer = crayfish. • Descriptive knowledge: • 1848 - increased incidence of breast cancer in nuns (childlessness, non-breastfeeding). • 1902 - Connection of X-rays and cancer induction . • 20th century - occurrence of familiar type of tumors. 21 Department of Experimental Biology MUNI SCI Classification of tumors • Based on their ability to invade other tissues. • Benign - they remain at the place of their origin, do not migrate, do not invade other tissues. • Malignant - they invade into surrounding tissues and, through the blood and lymphatic system, throughout the body, in new tissues they provoke the formation of secondary tumors (metastases). • From this point of view, tumors can be classified into primary and secondary as well. 22 Department of Experimental Biology @Jana Šmardové MUNI SCI Classification of tumors • Based on their ability to invade other tissues. Benign Tumor Malignant Tumor and organs. very Benign Tumors Malignant Tumors • Small • Large Slow-growing • Fast-growing • Non-invasive • Invasive • Well-differentiated • Poorly-differentiated Stay localized • Metastasize • Stay where they are. • Infiltrate, fnvade, destroy • Can't invade or surrounding tissue. metastasize. • Then metastasize to other parts of body. https://diffzi.com/benign-tumor-vs-malignant-tumor/ IUI U 111 23 Department of Experimental Biology https://www.vei7wellhealth.com/what-does-malignant-and-benign-mean-514240 SCI Stages of tumor development Hyperplasia - is the stage where genetically altered or abnormal cells show uncontrolled and rapid growth. Dysplasia - stage of tumor development where overgrowing cells change their original form. It consists of more immature cells than mature. In situ cancer - represent neoplastic lesion where cells do not go into the process of maturation, lost their tissue identity and grow without regulation. Malignant tumor - overgrowing cells invade other areas by rupturing basal membrane. Metastases occur when cancer cells reach to the distant parts through lymphatic system and blood circulation. Genetically altered epithelial cell Hyperplasia • Cell divides more rapidly Dysplasia • Cells change form In situ cancer • Cells slay in one place Malignant tumor (cancer) • Cancer cells invade normal tissue and enter blood and lymph • Metastases form at distant sites Metastases Blood vessel Normal underlying connective or muscle tissue action of fl0lv 24 Department of Experimental Biology https://www.researchgate.net/deref/https%3A0/o2F°/o2Ftel. archives-ouvertes.fr%2Ftel-00912341 MUNI SCI Classification of tumors • Based on their origin - from which cell types and tissues. • Carcinomas - tumors of epithelial cells (about 90% of human tumors). • Sarcomas - solid tumors of connective tissues - muscles, bones, cartilage. • Leukemia and lymphomas - derived from hematopoietic cells and cells of the immune system. • Gliomas - tumors derived from nervous tissue. MUNI 25 Department of Experimental Biology r> r» t Classification of tumors • Based on the invading organ. • lung cancer • colorectal cancer • breast tumor • acute myeloid leukemia • and many other types 26 Department of Experimental Biology MUNI SCI Cancerogenesis • Cancerogenesis or Carcinogenesis - is the process of tumor formation and development. It could also defined as the process of tumor onset and progression. • The essence of carcinogenesis is the gradual accumulation of genetic (and epigenetic) changes - mutations. • It is multistage process. • Neoplastic transformation - is the transformation of a somatic cell into a tumor cell. 27 Department of Experimental Biology MUNI SCI Cancerogenesis • Carcinogenesis - accumulation of genetic mutations and epigenetic) changes. • Under selection pressure, earlier diver mutations, and also passenger mutations obtain advantage for outgrowth of clones and drive tumors grow. C tonal Genetic _ Selection expansion alterations pressure OOO 09Ö Cancer cell Driver mutations 0 Passenger mutation MUNI 28 Department of Experimental Biology International Journal of Biological Sciences, July 2017, 13(8), 949-960. O 0 J. Cancerogenesis • The clonal expansion of a population of mutated cells (cancer cells) from an individual single-cell causes tumor heterogeneity in pathology and molecular profiles. • Tumor heterogeneity within tumors is created by genetic and epigenetic changes. V-1 clone 2MlClone 3rd Clone Increase mutations 29 Department of Experimental Biology Cancers, April 2020, 12(1050) MUNI SCI Cancerogenesis • Nowell's theory of tumor evolution. • The tumor arises from a single cell. • Dominance of one clone. • Aggressive clones spread, passive ones perish • Selection, clonal expansion. • Clonal diversity is accompanied by genetic heterogeneity. Primary tumor Metastasis 1 O 0 00000 Normal Founder Subclones with cell clone individual genotypes 30 Department of Experimental Biology Nature Biotechnology volume 30, pages408-410 (2012) Science. 1976 Oct 1 ;194(4260):23-8. Blood. 2012 Aug 2;120(5):927-8. MU NI SCI Cancer multistage process • Colonic epithelial cells undergo a histologic transition from normal to malignant state that is driven by specific genetic events including inactivation of tumor suppressors (APC, SMAD4 and TP53) and activation of the KRAS oncogene. • The three stages of adenomas represent tumors of increasing size, dysplasia, and villous content. APC KRAS (50%) SMAD4 TP53 ( \ I r-> r ■> v ( —' r > Normal t Aberrant Early ^ 1 Intermediate Late Carcinoma Invasion. epithelium 1 ^--/ crypt foci adenoma I adenoma adenoma m situ Mm metastasis 31 Department of Experimental Biology November 201 OSmall GTPases 1(1):2-27 MUNI SCI Tumor - histology • All tumors, benign and malignant, have two basic components: • Stroma - supporting, host-derived, non-neoplastic, made up of connective tissue, blood vessels, and host-derived inflammatory cells. o Crucial to the growth of the neoplasm, o Carries the blood supply. o Provides support for the growth of parenchymal cells. • Parenchyma - made up of transformed or neoplastic cells, o Largely determines its biologic behavior. o Tumor derives its name. 32 Department of Experimental Biology https://www.slideshare.net/DeepakKumarGupta2/general-aspect-of-neoplasia MUNI SCI Tumor is a complex tissue • The interactions between the genetically altered malignant cells and these supporting coconspirators proves to be critical in cancer onset and progression - microenvironment. The Reductionist View A Heterotypic Cell Biology 33 Department of Experimental Biology https://www.nature.com/subjects/double-strand-dna-breaks MUNI SCI • The Hallmarks of Cancer 34 Department of Experimental Biology The hallmarks of cancer Hanahan and Weinberg Avast SafeZone v * Přihlášení The hallmarks of cancer. - X + ♦ S ■■■ A www.ncbi.n[m.rihj.gav^pubmed/'ir>647931 + ! Chcete-li mít vase záložky stále pa ruce, přidejte je na tutá listu řj NCBI ResoNicesQ HmiToS PublÄJcd;.. I PubMed S3 Format Abstract - Cell 2O0(> Jan 7,1 W)[1>:57-7D. The hallmarks of cancer. Hanahan □'. ■'-sinl.'^rc R-. h Author information 1 Department of Biochemistry. Hormone Research Institute. University of California at San Francisco. 94143. USA. PMID: 1064793* [Ind&xetf for MEDLINE] Free full tent 19 872 citations Publication types, MeSH terms LinkOut - more resources Full text links I Cell I Save items m ů Add to Favorites H Similar articles Cancer research. Obstacle for promising cancer therapy. [Science. 2002] Is oncogene addiction angicgenesis-dependent? [Cold Spring Harb Symp Quant Bi.__] Angiogenesis. Successful growth of tumours [Nature. 1989] [Apoptosis or programmed cell death] [Ann Pathol. 19%] 3 [Studies of growth differentiation and neoplastic transfornialii [Postepy Biochem 1938] See reviews... See ell... 35 Department of Experimental Biology Cell. 2000 Jan 7;100(1):57-70. MUNI SCI The hallmarks of cancer • Robert Weinberg • 1982 - Discovered Ras, the first human oncogene • isolated Rb for the first time in his lab •MIT • Dough Hanahan •MIT, UCSF • Director of Ecole Polytechnique Federale de Lausanne • 1983- SOB medium for bacteria Robert Weinberg 36 Department of Experimental Biology Douglas Hanahan |\/| U |\| SCI What genes are altered in cancer? • A tumor is not a monogenic disease. • It is estimated that 4-7 events (interventions) are necessary for the development of the tumor. • There are dozens of specific genes that can be altered during carcinogenesis. • In general, there are six (seven?) basic acquired features of a fully malignant tumor. 37 Department of Experimental Biology https://encyclopedia.pub/entry/176 MUNI SCI Tumors must acquire the same hallmarks capabilities > Weinberg & Hanahan > Genome instability as a necessary condition for the accumulation of all necessary mutations? Component Acquired Capability Example of Mechanism n Self-sufficiency in growth signals Insensitivity to anti-growth signals Activate H-Ras oncogene Lose retinoblastoma suppressor n Evading apoptosis Limitless replicative potential Produce IGF survival factors Turn on telomerase in Sustained angiogenesis Produce VEGF inducer Li Tissue invasion & metastasis Inactivate E-cadherin 38 Department of Experimental Biology Cell. 2000 Jan 7;100(1):57-70. @Jana Šmardové MUNI SCI Tumors must acquire the same hallmarks capabilities Self-sufficiency in growth signals Evading 1 apoptosis 1 Insensitivity to anti-growth signals V Sustained 1 angiogenesis 1 Tissue invasion & metastasis Limitless replicative potential • Genome instability as a necessary condition for the accumulation of all necessary mutations? 39 Department of Experimental Biology Cell. 2000 Jan 7;100(1):57-70. MUNI SCI Cancerogenesis has an individual course • Thus, the order in which these capabilities are acquired seems likely be quite variable across the spectrum of cancer types and subtypes. • In some tumors, a particular genetic lesion may confer several capabilities simultaneously, decreasing the number of distinct mutational steps required to complete tumorigenesis. -x 40 Department of Experimental Biology Cell. 2000 Jan 7;100(1):57-70. Tumors must acquire the six hallmarks capabilities Component Acquired Capability Example of Mechanism 41 Department of Experimental Biology MUNI SCI Sustainig cell signaling • Healthy cells - carefully control growth and division. The signal, binding to receptor, triggering of the signaling pathway. • Tumor cells deregulate these signals, they do not need stimulation by growth factors to proliferate. • Tumor cells can produce: o Their own growth factors, such as PDGF - glioblastomas, o Receptor overexpression - EGFR - breast and stomach tumors, o Change the receptor structure - receptor does not need a ligand to activate. 42 Department of Experimental Biology MUNI SCI Growth factors and Receptor-tyrosine kinases • Growth factors: Polypeptides, which are produced by cells and induce signaling to start or stop proliferation, differentiation, survival,... by activating its specific receptors on the cell surface. • Stimulation: Three types of communication systems in biological systems differs manly by the distance that the communication operates over. o Autocrine - growth factor stimulates the producing cell, o Paracrine - stimulation of a neighboring cell, o Endocrine - stimulation of distant cells. Autocrine Paracrine Endocrine 43 Department of Experimental Biology http://www.labbookpages.co.uk/research/bioNode.html MUNI SCI Cell signaling This distribution is related to the structure of the signal pathway: Growth factor binding to receptor. Activation of receptor protein kinas. Signal transmission to the nucleus by a cascade of protein kinases. Activation of the transcription factor. Olher changes Protein kinase activity Adaptor proteins Docking proteins GTP-binding proteins Protein or lipid kinases Phosphodiesterases Metabolic enzymes Transcription factors Nuclear membrane gene expression 44 Department of Experimental Biology MUNI SCI Sustainig cell signaling • Cellular pathways critical for cell survival deregulated in tumors. Mutation - activation/inactivation of the pathway leads to: o Uncontrollable proliferation. o Invasiveness. o Resistance to signals. o Angiogenesis. o Metastasing. o Resistance to apoptosis. are 45 Department of Experimental Biology MUNI SCI Sustainig cell signaling EGFRJigands EGFR homo- o heterodimers Antibodies to EGFR STAT Cell Survival Cell proliferation Cell Migration • Activation of the EGFR pathway induces cellular survival and anti-apoptosis signals by activating transcription of genes associated with cell survival - NFKB and JAK/STAT. • EGFR is activated either as a homo- or heterodimer resulting in regulation of multiple pathways. • In particular the RAS/RAF/MAPK, AKT, and JAK/STAT pathways downstream of EGFR play integral roles in cell migration, proliferation, and survival, respectively. • Anti-EGFR antibodies are targeted to the external ligand binding domains while the small molecule inhibitors or tyrosine kinase inhibitors (TKIs) target its cytoplasmic kinase domains. 46 Department of Experimental Biology https://www.aacc.org/cln/articles/2013/october/egfr-mutations MUNI SCI Tumors must acquire the six hallmarks capabilities Component Acquired Capability Example of Mechanism 47 Department of Experimental Biology MUNI SCI Tumor as disease of the cell cycle • Loss of the cell cycle regulation is a critical part of cell transformation. • Loss of the cell cycle regulation is not the only part of cancerogenesis. • When altered it is not fully transformative itself. 48 Department of Experimental Biology MUNI SCI Cell cycle 49 Department of Experimental Biology https://quizizz.com/admin/quiz/606682e1825246001f488abd/dna-structure-function-and-replication-basics MUNI SCI Cell cycle • Each phase of the cycle is catalyzed by specific cyclin-dependent kinase associated with cyclin. • Cyklin-dependentni kinazy (CDK): o Phosphorylates its substrates. o Typically have a catalytic and regulatory subunit, activity always dependent on cyclin binding. • Cyklins: o Their level fluctuates depending on the phase of the cell cycle, o They activate the appropriate CDK and direct it to its substrates, then they are quickly degraded. 50 Department of Experimental Biology MUNI SCI Cell cycle Triggers cells to move jrom GO to Gl and from Gl Into 5 phase, prepares the cell for DNA replication in 5 phase. activates DNA replication inside the nucleus InS phase. promotes the ossembly of the mitotic spindle and other tasks in the cytoplasm to prepare for mitosis. G2/M-phase cyclin Cyclin B GZ/M-phase CDK Degradation \ I of S-phase ^ S-phase CDK G,Phase S Phase G,Phase Mil.,.,'. Q Degradation of G2/M-phase cyclin CKIs (INK4. Cip/Kip) G1 /S-phase cyclin Cyclin D/E G1/S-phase S-phase cyclin Cyclin A O o o Q Degradation of G1/S-phase cyclin Department of Experimental Biology https://mysciencesquad.weebly.com/ib-hl-16u5.html MUNI SCI Cell cycle • Cell cycle. • Interphase • Mitosis. 52 Department of Experimental Biology https://link.springer.eom/article/10.1007/s10637-009-9236-6/figures/1 MUNI SCI pRb a principal regulator of cycle • Retinoblastoma protein - pRB is non-phosphorylated state and blocks the passage through the G1 - S restriction point: o Interacts with TF family E2F. o Blocks their ability to transactivate their target genes -necessary for the S phase. • Phosphorylation of pRB leads to its ability to bind E2F and passage through the restriction point is thus possible. Regulation of passage through the restriction point = regulation of pRB phosphorylation. 53 Department of Experimental Biology MUNI SCI Regulation of pRb activity Non-phosphorylated RB Bound to E2F Phosphorylation oíRb w Protein factors for cell cycle progr ession Phosphorylation of Rb Release E2F E2F migrate to nucleus induce transcription Nucleus 54 Department of Experimental Biology Oncotarget, February 2019 10(12). MUNI SCI Regulation of pRb activity • The activity of pRB is positively affected: o Cyclins D (D1, D2 and D3) with CDK4/CDK6. o Cyclin E and CDK2 complexes. • Negatively affected by cellulat CDK/cyclin inhibitors: o Cip/Kip family proteins - p21WAF1 a p27KIP1. o INK4 family proteins- p15INK4B a p16INK4A. • Mitogenic signaling leads to increased expression of cyclins D and decreased levels of inhibitors. • Antimitogenic signaling leads to a decrease in the level of cyclins D and induction of the CDK/cyclin inhibitors. • Mutation in Rb - often lead to an uncontrollable division due to hyperphosphorylated status of pRb. 55 Department of Experimental Biology MUNI SCI pRb a principal regulator of cycle 56 Department of Experimental Biology Cancers 2021, 13(9), 2226. MUNI SCI Tumors must acquire the six hallmarks capabilities Component Acquired Capability Example of Mechanism Evading apoptosis Produce IGF survival factors 57 Department of Experimental Biology MUNI SCI Cell death and tumors progression OUTCOME A, líalanťťíl [írttlilf-riit km and ik'ulh • The rate of tissue growth is determined by the rate or equilibrium of cell division and the rate of cell death. • During homeostasis, both processes are in balance. • In a tumor, the balance between cell division and cell death is broken. 58 Department of Experimental Biology MUNI SCI Cell death and tumors progression Apoptosis: Cells live for a limited time. Then apoptosis (programmed cell death) is triggered. Tumor cells are resistant to the induction of cell death due to incapability to respond to death signals or they become sensitive to the oncogenic signals. Growth-promoting pathways Oncogenic s^r* changes Proliferation Cell survival Intrinsic tumor suppressor pathways i i \_i. 59 Define footer - presentation title / department Future Aspects of Tumor Suppressor Genehttp://dx.doi.org/10.5772/56471 MUNI SCI Necrosis Swelling of endoplasmic reticulum and mitochondria Membrane blebs Breakdown of plasma membrane, organelles and nucleus; leakage of contents NECROSIS Amorphous densities in mitochondria Copyright £ 2010 by SavnctT an mprint of Elsevier Inc. Necrosis refers to a group of affected cells caused by non-physiological damage (viral infection, hypothermia..), significant inflammatory reaction. Morphological features: o Begins with swelling of the cytoplasm and mitochondria organelles crumble, o Loss of cell membrane integrity. Biochemical features: 60 Department of Experimental Biology o Loss of regulation of homeostasis. o Passive process without energy (runs even in 4°C). o Random DNA degradation. MUNI SCI Apoptosis Apoplotic i^-'N body Phagocyte Phagocytosis of apoptotic cells and fragments Copyright-c 2010 by Saunws, an mprint of Els*viw nc 61 Department of Experimental Biology Apoptosis is programmed cell death: Affects individual cells. Induced by physiological stimuli (lack of growth factors) no inflammatory reaction physiological. Morphological features: o Condensation of the cell and nucleus, o „Blembs" of cell membranes, o Chromatin condensation, o Nucleus fragmentation and cell fragmatation into apoptotic bodies. • Biochemical features: Strictly regulated, active process. o MUNI SCI Regulation of apoptosis 62 Department of Experimental Biology External receptor pathway - ligand binding to the corresponding death receptor produces a proapoptotic signal. This leads to the activation of the domain of death with the cooperation of other proteins. The goal is to activate procaspase 8. Internal path - signaling e.g. through p53 initiates an apoptotic process, the component of which is the Bcl-2/Bax system, signaling through mitochondria, the formation of a complex called apoptosome and the goal is to activate procapspase 9. MUNI SCI Dysregulation of apoptosis in cancerogenesis Apoptosis is dysregulated in cancer leading to disruptions of the delicate balance between cell proliferation and cell death. Examples of altered apoptotic proteins in tumors: o Up-regulation of Bcl-2 (chromosomal translocation) in lymphomes. o Up-regulation of survival factors IGF-1, IGF-2. o Mutation and downregulation of the Fas death receptors. o Mutations. o Actiovation of bax. o Inactivation of p53. 63 Department of Experimental Biology Cancers , 2019, 11(11), 1623. Defective death signaling Reduced expression of death receptors and ligands -J?rf receptors \\— Truncated death receptors lacking intracellular domains Mutated death receptors Pathways: PI3K Notch Hedgehog IMF-kB Hippo-YapJ ^ f Imbalanced ratio of pro-apoptotic to anti-apoptotic proteins MUNI SCI Tumors must acquire the six hallmarks capabilities Component Acquired Capability Example of Mechanism Limitless replicative potential Turn on telomerase 64 Department of Experimental Biology MUNI SCI Replication potential • Mammalian cells have a replication potential of 60-70 divisions (Hayflick limit). • Then they go to the stage of senescence - they change the morphology, metabolically active, do not divide, block CC - p53, Rb. • Mutation in p53 or Rb - another 30 divisions - crisis, chromosomal aberration - apoptosis. • If the mutation occurs with probability 10-7 - then there is a great chance that immortal cell will appear. • Most tumor cells - immortal. MUNI 65 Department of Experimental Biology r> r» t Telomerase hypothesis When the critical length of the telomeres is reached, signals are triggered that induce a state of senescence. With further division (inactivation of p53 and pRB), telomeres are further shortened and cause chromosomal instability, which provokes a crisis. Telomere shortening acts as a mitotic counter that determines the proliferative capacity of all cell types that do not have telomerase activity. . parental strand 3' TELOMERASE BINDS 3TTGGGGTTGGGGTTGGGGTTG -^AACCC5'X incomplete, newly synthesized lagging strand 3' 3TTGGGGTTGGGGTTGGGGTTG 3AACCCC MACCCCAAC TELOMERASE EXTENDS 3' END (RNA-tem plated DNA synthesis) 5' é direction of telomere synthesis telomerase with bound RNA template ITTGGGGTTGGGGTTGGGGTTGGGGTTGGGGTTG ÜAACCCC ^ACCCCAAC 5' COMPLETION OF LAGGING STRAND BY DNA POLYMERASE (DNA-templated DNA synthesis) (f 3' 3' ]TTGGGGTTGGGGTTGGGGTTGGGGTTGGGGTTG ]AACCCC .CCCCAACCCCAACCCC 5' DNA polymerase 66 Department of Experimental Biology Clear view of telomerase at last (acs.org') https://cen.acs.org/biological-chemistry/genomics/Clear-view-telomerase-last/96/i18 MUNI SCI Telomerase • Germ cells. • Some stem cells. • Some somatic cells under specific conditions: • Lymphocytes activated by mitogens. • Cells in the proliferative zone of intestinal crypts. • Cells of the proliferative basal layer of the skin. • Cells of the lobular endothelium of the breast during pregnancy. 67 Department of Experimental Biology MUNI SCI Telomerase hypothesis Normal dividing cell Multiple replications, progressive telomere shortening Abrogation of p53 and/or pi 6/Rb tumor suppressor pathways Extended lifespan Loss of cell cycle checkpoint control Crisis - M2 phase Uncapped telomeres Chromosome end-to-end fusions Incipient immortal cell High genomic instability Threshold short telomere length Cellular senescence M1 phase DNA damage activation Cell death Dysfunctional telomeres Extensive cell death due to mitotic catastrophe hTERTor ALT activation Immortal cancer eel Telomere shortening is a natural consequence of cell division due to the "end replication problem". In the cells undergoing replicative senescence, the p53 and p16-pRB pathways are often activated leading to essentially irreversible growth arrest. Cells that gain additional oncogenic changes (p53 loss) can bypass senescence and continue to divide until chromosome end-to-end fusions. Only a rare human cell (one in 105 to 107) can engage a mechanism to bypass crisis and become immortal. This is almost universally accomplished by the upregulation or reactivation of telomerase. A rarer telomerase negative immortalization pathway, termed ALT (alternative lengthening of telomeres), involves DNA recombination to maintain telomeres. 68 Department of Experimental Biology Genome Med. 2016; 8: 69. MUNI SCI Tumors must acquire the six hallmarks capabilities Component Acquired Capability Example of Mechanism Sustained angiogenesis Produce VEGF inducer 69 Department of Experimental Biology MUNI SCI Angiogenesis in cancerogenesis Department of Experimental Biology Tumor-associated Angiogenesis Growth of blood capillaries from the existing vasculature. An important step - from the dormant state of the tumor to the "^gP^S^S10" malignant. Tumor - population of rapidly and uncontrollably growing cells. Tumors can not grow more than 1 - 2 mm3 several million cells (lack of nutrients and oxygen). HIF-1 activates vascular endothelial growth factor - VEGF. Without angiogenesis, the tumor grows slowly and linearly, then exponentially. Stimulation of angiogenesis by VEGF 42r Rapid tumor growth and metastasis Molecular Cancer Therapeutics Reviews MUNI SCI VEGF NRP-1 IMRP-2 Co-receptors VEGF-B PLGF VEGF-A VEGF-C VEGF-0 VEGFR-1 (Flt-1) VEGFR-2 (KDR/Flk-1) VEGFR-3 (Flt-4) VEGFR-1/-2 VEGFR-2/-3 heterodimer heterodimei' 1 Angiogenesis (developrnenlal) Angiogenesis Proliferation Migration Invasion Survival Vascular permeability Lymph-angiogenesis • VEGF was the first characterized factor specific to vasculogenesis - vascular endothelial growth factor (original name - vascular permeable factor). • It is critical for the initiation of vasculorogenesis as well as for angiogenic branching. • Today described 5 different VEGF factors and 3 different receptors. • Interactions between VEGF and VEGF receptors orchestrate distinct biological functions. 71 Department of Experimental Biology Biomolecules and Therapeutics, January 2015, 23(1):12-8. MUNI SCI Tumor angiogenesis Tumor angiogenesis: (A) Tumor cells produce VEGF-A and other angiogenic factors such as bFGF and angiopoietins. These stimulate endothelial cells to proliferate and migrate. (B) An additional source of angiogenic factors is the stroma. This is a heterogeneous compartment, comprising fibroblastic, inflammatory, and immune cells. (C) Endothelial cells produce PDGF-ß, which promotes recruitment of pericytes in the microvasculature after activation of PDGFR-ß. 72 Department of Experimental Biology OncoTargets and Therapy, December 2014, 7(default):2237-2248 MUNI SCI Tumor angiogenesis • Tumor vasculature is highly disorganized. • This can arise as a result of the uneven release of angiogenic regulators. • The flow of blood is chaotic in different parts of the system. • Therefore, places with hypoxia and excessive acidity are formed in the tumor. • This circumstance may affect the effect of therapy; a space is created in which, for example, the selection and clonal expansion of cells that do not respond to hypoxia by apoptosis. Control +VEGF inhibitor • Microcomputed tomography image showing effect of VEGF inhibition in a preclinical model. 73 Department of Experimental Biology https://www.genentechoncology.com/pathways/cancer-tumor-targets/vegf.html MUNI SCI Tumors must acquire the six hallmarks capabilities Component Acquired Capability Example of Mechanism Tissue invasion & metastasis Inactivate E-cadherin 74 Department of Experimental Biology MUNI SCI Tumors progression Primary tumor formation Extravasation Micrometastasis formation ■4Ž 75 Department of Experimental Biology Local invasion Arrest at a distant organ site Metastatic colonization Intravasation Survival in the circulation Clinically detectable macroscopic metastases MUNI SCI Metastatic cascade Primary Tumor Site Blood Stream Step 3: Circulation 76 Department of Experimental Biology Metastatic Site ® ® Tumor Cell Endothelial Cell Stromal Cell © Inflammatory Cell ECM Step 5: Colonization \ Step 4: Extravasation • Overview of the Metastatic Cascade. • Step 1: Invade through basement membrane and migration. • Step 2: Intravasation into vasculature. • Step 3: Circulation of tumor cells in the bloodstream to blood vessels around secondary sites. ° )C_1 • Step 4: Extravasation through the J^o) endothelial barrier. • Step 5: Colonization in the " ~~ metastatic target organ. npj Precision Oncology, 2019, volume 3, Article number: 20. MUNI SCI The mechanism of metastasis Wnt molecule Ml.....Ulili. ■ Mill III I i ■ i M UJMI Uli IIJ [| 1 1''1''1 'i'l nucleus C-Myc * Cell migration Cell proliferation Cell growth * (Apoptosis) • Adhesive molecules - N-CAM - adhesive molecule, increased expression in Wilms tumor, neuroblastoma. • E-cadherin - on epithelial cells, antiproliferative signals, tumors reduce expression. • Integrins - changes in expression on migrating cells. 77 Department of Experimental Biology Journal of Clinical Pathology, August 2013, 66(11) MUNI SCI The mechanism of metastasis Wnt TGF-ß • Glycoprotein important for cell adhesion. • Differentiation of epithelial cells. • Loss of expression - epithelialmesenchymal transition, an important step in the metastatic progression of human tumors. • Tumor suppressor. • Decreased expression in epithelial tumors - invasiveness and worse prognosis. 78 Department of Experimental Biology Int. J. Mol. Sei. 2020, 21, 7624; doi:10.3390/ijms21207624 MUNI SCI • The Hallmarks of cancer: The next generation MUNI 79 Department of Experimental Biology _ _ T O 0 J. Hallmarks of cancer: The next generation Hanahan and Weinberg O R Hailmarks of cancer: the next ge X G Google ■■■■^■■■■■■■■■■■■bH & O to) Ů https://pubrned.nebi.hlra nih.gov/21376230/ (5 Bakaláři - mezi škol... Informační systém Q Amazon.co.uk- On... Q Booking.com 80 Department of Experimental Biology National Library of Medicine National Center for Biotechnology Information Pubty^ed gov Winberg Hanahan 2011| Advanced Create alert Create RSS Found 1 result for an alternative search. Your search for Winberg Hanahan 2011 retrieved no results. Review > Cell. 2011 n Mar^^e ^^J^e/j^jrjiQil j Q IH S Hallmarks of cancer: the next generation Douglas Hanahan 1, Robert A Weinberg Affiliations +• expand PMID: 21376230 DOI: 10.1016/j.cell.2011.02.013 Free article Abstract + O o x Další" cbnbené položky Save Email Send to Display options -jjt- FULL TEXT .INKS :.uj;iMri=i.-!j ti Cite É Favorites The hallmarks of cancer comprise six biological capabilities acquired during the multistep development of human tumors. The hallmarks constitute an organizing principle for rationalizing the complexities of neoplastic disease. They include sustaining proliferative signaling, evading growth suppressors, resisting cell death, enabling replicative immortality, inducing angiogenesis, and acttvatinq invasion and metastasis. Underlying these hallmarks are qenome instability, which :: p l. o & d ■ - * D «r Cell, Volume 144, Issue 5, 4 March 2011, Pages 646-674 O O O PAGE NAVIGATION MUNI SCI Tumors must acquire additional hallmarks capabilities Deregulating cellular energetics Genome Instability and mutation ^Emerging Hallmarks^- Avoiding Immune destruction Tumor-promoting Inflammation Deregulating cellular energetics - modification or reprograming of cellular metabolism in order to effectively support neoplastic proliferation. Avoiding immune destruction - allows cancer cells to evade immunological destruction mediated by T and B lymphocytes, macrophages, and natural killer cells. Genomic instability and mutation - endow cancer cells with genetic alterations that drive tumor progression. -(^Enabling Characferistics^)- Tumor-promoting inflamation - inadvertent support of multiple hallmark capabilities due to inflammatory responses of innate immune cells. 81 Department of Experimental Biology Cell, Volume 144, Issue 5, 4 March 2011, Pages 646-674 MUNI SCI Tumors must acquire additional four hallmarks capabilities -^Emerging Hallmark$y Deregulating cellular energetics Genome Instability and mutatfon At/elding Immune destruction Tumor-promoting Inflammation 82 Department of Experimental Biology ^Enabling Characteristics^) Cell, Volume 144, Issue 5, 4 March 2011, Pages 646-674 MUNI SCI Genome instability and mutation Genomic instability and mutation - endow cancer cells with genetic alterations that drive tumor progression. This may be acquired through... o Clonal selection - through nonmutational changes affecting the regulation of gene expression. o Epigenetic mechanism - DNA methylation and Histone modifications. Alterations in DNA maintenance machinery due to defects in proteins involved in: Detecting DNA damage and activating the repair machinery. Directly repairing damaged DNA. Inactivating or intercepting mutagenic molecules before they have damaged the DNA. MUNI Department of Experimental Biology r> r» t Genome instability and mutation Potential mechanisms of action Inhibition of PARP enzyme activity Single-strand breaks PARP trapping Persistent unrepaired single-strand breaks T 1 PAR Pi Synthetic lethality in HR deficient tumours (eg BRCAm tumours) Trapping of PARP-DNA complexes Lethality potentially not restricted to HR deficiency (^J) PARP H PARP inhibitor ^ Poly ADP ribosylation DNA damage Inactivating or intercepting mutagenic molecules before they have damaged the DNA. Inhibition of PARP enzyme activity or catalytic inhibition interferes with the repair of single-strand breaks, leading to stalled DNA replication forks that requires HR repair. In HR-deficient tumors, such as those with BRCA mutations, PARP inhibition results in synthetic lethality. PARP trapping refers to trapping of PARP proteins on DNA, which also leads to replication fork damage, but because this pathway utilizes additional repair mechanisms, it is not restricted to tumors with HR deficiency. 84 Department of Experimental Biology British Journal of Cancer volume 119, pages141-152 (2018) MUNI SCI Tumors must acquire additional four hallmarks capabilities -^Emerging Halltnark$y Deregulating cellular energetics Avoiding Immune destruction Genome Instability and mutatfon 85 Department of Experimental Biology ^Enabling Characteristics^) Cell, Volume 144, Issue 5, 4 March 2011, Pages 646-674 MUNI SCI Tumor promoting inflammation • Every neoplastic lesion contains immune cells present at densities ranging from subtle infiltrations detectable only with cell type-specific antibodies to gross inflammations. • Inflammation contributes to multiple hallmark capabilities by supplying bioactive molecules to the tumor microenvironment, including: o Growth factors that sustain proliferative signaling, o Survival factors that limit cell death, o Proangiogenic factors. o Extracellular matrix-modifying enzymes that facilitate angiogenesis, invasion, and metastasis, o Signals that lead to activation of EMT. o Various factors facilitating other hallmarks programs. 86 Department of Experimental Biology MUNI SCI Tumor promoting inflammation • Most solid tumors are assembled of distinct cell types. • Both the parenchyma and stroma of tumors contain distinct cell types and subtypes that collectively enable tumor growth and progression. •Also, the immune inflammatory cells present in tumors can include both tumor-promoting as well as tumor-killing subclasses. 87 Department of Experimental Biology Cancer-Associated Fibroblast (CAF) Endothelial Cell (EC) Pericyte (PC) Local & Bone marrow-derived Stromal Stem & Progenitor Cells Cancer Stem Cell (CSC) Cancer Cell (CC) CO Immune Inflammatory Cells (ICs) L Invasive Cancer Cell MUNI SCI Tumor promoting inflammation PD-L1 binds to PD-1 and inhibits T cell killing of tumor cell Tumor cell PD-L1 Antigen T cell receptor PD-1 cell 88 Department of Experimental Biology Blocking PD-L1 or PD-1 allows T cell killing of tumor cell Tumor cell death PD-L1 cell 2015 Terase Winstow LLC S. Govt, has certain rights • Checkpoint proteins, such as PD-L1 on tumor cells and PD-1 on T cells, help keep immune responses in check. • The binding of PD-L1 to PD-1 keeps T cells from killing tumor cells in the body. • Blocking the binding of PD-L1 to PD-1 with an immune checkpoint inhibitor (anti-PD-L1 or anti-PD-1) allows the T cells to kill tumor cells. https://www.cancer.gov/about-cancer/treatment/types/immunotherapy/checkpoint-inhibitors MUNI SCI Tumor promoting inflammation • T Cells have CTLA-4 that works as a regulating switch for T cell activity. • By blocking the CTLA-4 pathway that T cell activation can be restored. • Ipilimumab is a drug blocking the CTLA-4 protein. When this protein is blocked the T-cell is activated and the tumor cell is killed. CTLA-4/B7 binding inhibits Blocking CTLA-4 allows T cell activation T cell killing of tumor cell 89 Department of Experimental Biology https://www.biologycorner.com/2020/05/27/jim-allison-breakthrough/ MUNI SCI Tumors must acquire additional four hallmarks capabilities -^Emerging Halltnark$y Deregulating cellular energetics Gencme Instability and mutatfon Auoidinů Immune destruction Tumor-promoting Inflammation 90 Department of Experimental Biology ^Enabling Charactefistics^)- Cell, Volume 144, Issue 5, 4 March 2011, Pages 646-674 MUNI SCI Deregulating cellular energetics • Reprogramming energy metabolism. • The Warburg Effect: Cancer cells reprogram their glucose metabolism, by limiting their energy production largely to glycolysis. • The cells upregulate glucose transporters, notably GLUT1, which substantially increases glucose import into the cytoplasm. • Growth factor-independent activation of the PI3K/Akt and c-Myc pathways facilitates increased rates of glucose uptake and glycolysis. • The hypoxia response system acts pleiotropically to upregulate glucose transporters and multiple enzymes of the glycolytic pathway. 91 Department of Experimental Biology Genes & Dev. 2009. 23: 537-548 MUNI SCI Deregulating cellular energetics Glc Growth Factor AAs Lac 92 Department of Experimental Biology Genes & Dev. 2009. 23: 537-548 Deregulating cellular energetics Glc t Glc Hißtet J \ Pyr PDK1 —I PDH Fatty Acids Bioenergetic Stress Lipids LKB1 I TSC1-TSC2 AAs I / 1 I ACC1 |— AMPK—I TORC1-«— Rag t T Bax_i Bcl2 Bak 'bcIXl Angiogenesis Autophagy Beclin-11— Bcl2 BclX, Apoptosis HIF1« degradation vhl q HIF1a Glycolysis 93 Department of Experimental Biology Genes & Dev. 2009. 23: 537-548 • Intracellular sensors of energy, nutrients, and oxygen promote metabolic adaptation to stress during tumorigenesis. • Major physiological strategies of metabolic adaptation include: oCell cycle inhibition. o Inhibition of biosynthetic pathways (lipid, protein synthesis), o Increases in bioenergetic pathways (p-oxidation, glycolysis, and OXPHOS). o Induction of autophagy. • Oncogenes are displayed in green and tumor suppressors in red. MUNI SCI Tumors must acquire additional four hallmarks capabilities -^Emerging Hallmarks^ Deregulating cellular energetics Genome Instability and mutatfon Tumor-promoting Inflammation 94 Department of Experimental Biology ^Enabling Characteristics^)- Cell, Volume 144, Issue 5, 4 March 2011, Pages 646-674 MUNI SCI Evading immune destruction • Cells and tissues are under constant surveillance by the immune system and immune surveillance is responsible for recognizing and eliminating the vast majority of incipient cancer cells and thus nascent tumors. • Tumors somehow managed to avoid detection by the various arms of the immune system or have been able to limit the extent of immunological killing, thereby evading eradication. • Immune system operates as a significant barrier to tumor formation and progression, at least in some forms of non-virus-induced cancer. • Mice lacking NK and T cells were more susceptible to cancer development. • Patients with higher CTLs and NK cells have a better prognosis. 95 Department of Experimental Biology MUNI SCI Evading immune destruction Nascent transformed cells are directly eradicated by innate and adaptive immune responses. During tumor growth, tumor cells are required for angiogenesis and stromal remodeling, which produce tumor cell variants that have low immunogenicity and are resistant to immune attack. Tumor progression leads to the release of tumor-derived soluble factors that are involved in several mechanisms of immune evasion in the escape phase. ElimenaLKjn rumom immure sujrwinsttít Equilibrium Escape TDSFs Immunological i^noiance 11 '^H; ■0 BM (il N Immunological tolfiianfis- 96 Department of Experimental Biology • iDC, immature dendritic cell; TAs, tumour antigens; SLN, sentinel lymph node; TAM, tumour-associated macrophage; TDSFs, tumour-derived soluble factors; Tregs, regulatory T cells; BM, bone marrow. II II l\l T mice with combined immunodeficiencies in both T cells and NK cells were even more ll/l U 111 1 susceptible to cancer development SCI Evading immune destruction Progression to metastatic disease is generally accompanied by decreased or altered TGF-p responsiveness and increased expression or activation of the TGF-p ligand. Normal epithelium Stromal microenvironment e. g. inflammation Changes in genetic and epigenedc context ——r^^^-^^^^ ^^^^ ^^^^ Invasive metastatic cancer Suppressor activities dominate Tumor ceil autonomous •growth inhibition ■ a po ptosis 'genomic stability Pro-oncogen ic activities dominate Tumor ceil autonomous 'ETAT •invation/motality • survival Effects on tumor stroma • immunosuppression •angiogenesis Tumor microenvironment 97 Department of Experimental Biology Trends in Immunology, Volume 31, Issue 6, June 2010, Pages 220-227 MUNI SCI Evading immune destruction • TGF-p affects multiple components of the immune system. • TGF-p inhibits the function of natural killer (NK) and CD8+ CTL (cytotoxic T lymphocytes), by blockinproduction of perforin, granzymes and cytotoxins. • TGF-p induces Treg and Th17 cell differentiation and inhibits B-cell proliferation and dendritic function. • TGF-p, inhibits macrophage and neutrophil development, but promotes type II macrophages and neutrophils, and mediates the immune suppression function of MISCs. F i*p3 Ftfrioflru gianzyme A and TGFfí regulatifln of immune calls in tumor microcnvirc-nment tm iWSiv? -f i ví: :."v, 98 Department of Experimental Biology Trends in Immunology, Volume 31 , Issue 6, June 2010, Pages 220-227. MUNI SCI Therapeutic Targeting of the Hallmarks of Cancer CEGFR A inhibitors J (Cyclin-dependentN kinase inhibitors J Aerobic glycolysis inhibitors CProapoptotic A ^ BH3 mimetics J** Deregulating cellular energetics Resisting cell dealh Sustaining proliferative signaling Evading growth suppressors Immune activating anti-CTLA4 mAb PARP inhibitors Genome instability & mutation 99 Department of Experimental Biology Cell, Volume 144, Issue 5, 4 March 2011, Pages 646-674 MUNI SCI • The Hallmarks of Cancer: New Dimensions MUNI 100 Department of Experimental Biology _ _ T O U J. Hallmarks of cancer: New Dimensions In 2022 additional emerging hallmarks and enabling characteristics of cancer were proposed: Unlocking phenotypic plasticity. Nonmutational epigenetic reprogramming. Polymorphic microbiomes. Senescent cells. Unlocking phenotypic plasticity Emerging hallmarks & enabling characteristics Nonmutational epigenetic reprogramming Senescent cells Polymorphic microbiomes 101 Department of Experimental Biology Cancer Discov (2022) 12 (1): 31-46. MUNI SCI Tumors acquire additional four hallmarks capabilities 102 Department of Experimental Biology MUNI SCI Phenotypic plasticity Phenotypic plasticity enables various disruptions of cellular differentiation, including: Dedifferentiate from mature to progenitor states. Blocked (terminal) differentiation from progenitor cell states. Transdifferentiation into different cell lineages. Progenitor cell II mi 111 Differentiated cell Normal differentiation 3* * Dedifferentiation Blocked differentiation Transdifferentiation 103 Department of Experimental Biology MUNI SCI Phenotypic plasticity For example the developmental transcription factors HOXA5 are highly expressed in differentiating colonic epithelial cells, and typically lost in advanced colon carcinomas, which characteristically express markers of stem and progenitor cells. 104 Department of Experimental Biology Cancer Cell, Volume 28, Issue 6, 14 December 2015, Pages 683-685 MUNI SCI Phenotypic plasticity Cancer Stem Cell Differentiated Cell WNT A LDH': 3 APCDD1 CXKC4 NKD1 stem less differentiation In colon cancer, HOXA5 is down regulated, and its re-expression induces loss of the cancer stem cell phenotype, preventing tumor progression and metastasis. Tumor regression by HOXA5 induction can be triggered by retinoids, which represent tangible means to treat colon cancer by eliminating cancer stem cells. MUNI 105 Department of Experimental Biology CancerCell, Volume 28, Issue 6, 14 December2015, Pages 815-829 O 0 J. Tumors acquire additional four hallmarks capabilities 106 Department of Experimental Biology MUNI SCI Nonmutational epigenetic reprogramming Nonmutational epigenetic regulation of gene expression is of course well established as the central mechanism mediating embryonic development, differentiation, and organogenesis. Analogous epigenetic alterations can contribute to the acquisition of hallmark capabilities during tumor development and malignant progression. 5 "J Epithelial to Mesenchymal Transition Tumor Epithelial cells Early hybrid EMT cells Late hybrid EMT cells Tumor Mesenchymal cells E > O E > TJ O C Ír TJ O E > 107 Department of Experimental Biology Cell Stem Cell, Volume 24, Issue 1, 3 January 2019, Pages 65-78 MUNI SCI Nonmutational epigenetic reprogramming normoxia hypoxia CG-CG—CG-CG—CG +aKG, Fe2+, VitC —CG—CG-CG-CG CG-CG—CG-CG" CG-CG—CG-CG-CG DNA demethylation CG-CG—CG-CG-CG DNA hypermethylation One common characteristic of tumors (or regions within tumors) is hypoxia, consequent to insufficient vascularization. Hypoxia, for example, reduces the activity of the TET demethylases, resulting in substantive changes in the methylome, in particular hypermethylation. Under hypoxic conditions (right) TET activity is compromised, leading to accumulation of methyl groups and causing DNA hypermethylation. 108 Department of Experimental Biology Mol Cell Oncol 2016;3:e1240549. MUNI SCI Tumors acquire additional four hallmarks capabilities 109 Department of Experimental Biology MUNI SCI Polymorphic microbiomes • Microbiota, that symbiotically associate with the barrier tissues of the body exposed to the external environment - the epidermis and the internal mucosa, in particular the gastrointestinal tract, as well as the lung, the breast, and the urogenital system. • For cancer, the evidence is increasingly compelling that polymorphic variability in the microbiomes between individuals in a population can have a profound impact on cancer phenotypes. • Microbiotic organisms have protective or deleterious effects on cancer development, malignant progression, and response to therapy. 110 Department of Experimental Biology MUNI SCI Polymorphic microbiomes Polymorphic microbiomes in one individual versus another, can diversely influence—by either inducing or inhibiting— many of the hallmark capabilities. Tumor microbiome, are implicated in modulating the acquisition - both positively and negatively - of the illustrated hallmark capabilities in certain tumor types. Gut Skin Lung Vaginal/ cervical Oral Tumor > Modulating tumor • Growth • Inflammation • Immune evasions ■ Genome instability ■Therapy resistance 111 Department of Experimental Biology MUNI SCI Polymorphic microbiomes Intact intestinal epithelial cells Dendritic cell (DC) Tolerant immune response I Decreased 1 inflammation Disrupted intestinal epithelial ^} 11 j| ^) cells Intestinal epithelial dysplasia and cancer Increased inflammation Dysregulated immune response ; 2017 American Association for Cancer Research Cancer Research Reviews AAG-R 112 Department of Experimental Biology Cancer Res (2017) 77 (8): 1783-1812. • Dysbiosis is immunocompromised state characterized by pathobiont colonization that leads to hyperinflammation, dysplasia, and tumorigenesis. • Pathobiont overgrowth leads to the loss of barrier integrity and a breach in the intestinal epithelial cell barrier. • The secretion of IL1 and IL6 from intestinal epithelial cells fuels a Th1 and Th17 response by DCs and macrophages and leads to higher levels of commensal-specific IgG by B cells. MUNI SCI Polymorphic microbiomes • Both anti-CTLA-4 and anti-PD-L1 therapies rely on gut microbiota for efficacy in immune activation. • Anti-PD-L1 therapy has been shown to rely on the preexistence of sufficient Bifidobacterium species, which are also thought to augment responses via PD-L1 binding on APCs, such as DCs and macrophages. • Similarly, anti-CTLA-4 indirectly alters the intestinal flora and enriches the Bacteroides species, possibly by promoting deterioration of the intestinal epithelial cell barrier via activation of local lymphocytes. Engages with flora i and T cells Promotes enrichment of resident t VN Bacteroides spp. in the gut jZe}~~~"~'*^ 1 \ 2017 American Association for Cancer Research Cancer Research Reviews AAG-R 113 Department of Experimental Biology Cancer Res (2017) 77 (8): 1783-1812. MUNI SCI Tumors acquire additional four hallmarks capabilities 114 Department of Experimental Biology MUNI SCI Senescent cells • Cellular senescence is a common outcome of various anticancer interventions. • Senescence-associated secretory phenotypes (SASPs) have pro-tumorigenic functions. • Evidence exists of increased cellular senescence in patients treated for various types of cancer. • Accumulation and persistence of therapy-induced senescent cells can promote tissue dysfunction and the early onset of various age-related symptoms in treated cancer patients. Immune cell recruitment Wound healing/ Fibrosis resolution Plasticity/Sternness Good Senescence reinforcement Cell fate/differentiation Tumor promotion Bad Paracrine Senescence Immune evasion Tissue dysfunction/ Inflammation 115 Department of Experimental Biology Trends in Cancer, Volume 6, Issue 10, October 2020, Pages 838-857 Genes & Dev. 2020. 34: 1565-1576 MUNI SCI Senescent cells HMGB1 SAA1 SAA2 Pro-inflammatory SASP I NF-kB KGATA4V 1 Nucleus (fEpigenetie regulation) ■ 7TG-p.p--— ~ ~/ " l r Jype 1 IFNs 'late SASP 1RF3U[ NF-kB u 116 Department of Experimental Biology Genes & Dev. 2020. 34: 1565-1576 • Regulation of the senescence-associated secretory phenotype (SASP). • Transcription factors - NF-kB, C/EBPß, STAT, MAML (yellow). • Intracellular signaling components - mTOR, TBK1, MAPK, JAK (orange). • Sensors and receptors and ligands - IL-6, IL-1,TLR2, NOTCH1 (red). MUNI SCI Senescent cells Senesecent col) K, SASP CCR2< myxoid nells NK cells Cancer ceil 117 Department of Experimental Biology Trends In Cnneor Trends in Cancer, Volume 6, Issue 10, October 2020, Pages 838-857 • Senescence-associated secretory phenotype (SASP) factors are engaged in senescent cells through the activation of the NF-kB, C/EBPp, and p38MAPK pathways. • SASP factors contribute to various aspects of cancer progression: • IL-6 and IL-8 promote EMT. • IL-6-IL-6R cancer cell proliferation. • CXCL5 and VEGF promote angiogenesis. • CXCL12-CXCR4 cancer cell invasion and migration. • IL-6 suppresses the CD45+CD3+ T cell-mediated immune clearance of cancer cells. • CCL2 suppresses the natural killer (NK). MUNI SCI Hallmarks of Cancer Sustaining proliferative signaling Evading growth suppressors Unlocking phenotypic plasticity Deregulating cellular metabolism Resisting cell death Genome instability & mutation Nonmutational epigenetic reprogramming Avoiding immune destruction Enabling repiicative immortality Tumor-promoting inflammation Senescent cells Polymorphic microbiomes Inducing or accessing vasculature Activating invasion & metastasis 118 Department of Experimental Biology MUNI SCI THANK YOU FOR YOUR ATTENTION "We're looking for somebody in medical research." 119 Department of Experimental Biology https://www.cartoonstock.eom/directory/l/lab_management.asp MUNI SCI