Cancers Identification of genes Function of genes Defining of the pathways Integration of the pathways Identification of Molecular Therapeutic Targets

Genes related with cancer; What genes are mutated in cancers ? Tumor suppressor, --suppress cell divisions; cell cycle controller Ex) G1/S, G2/M checkpoint controller Mutation(loss of function); uncontolled cell divisions & other mutations events Oncogene : promote the cell divisions Gain of functions; hyperactive Ex) Ras, EGFR active ?

Gatekeepers ( p53, Rb, ARF ets) Caretakers ( ATM, Brca1, PMS,MSH etc) Oncogene activation ( Ras, Src ets) Telomerase activation Mitogenic Signals DNA damage Signals Cell cycle arrest & Apoptosis Proliferations

The protein Networks

The Biology of Cancer First Edition Chapter 3: Tumor Viruses Copyright © Garland Science 2007 Robert A. Weinberg

Cancer is an infectious Disease ? No. –Virus-induced cancers represent only a minority of human cancers Study of tumor virus open many of the secrets of human cancer, even majority of cancer have no connection with tumor virus in human cancers. Studying tumor virus, identification of oncogenes, changed the direction of cancer research. 1876, Russian research reported that the transmission of a tumor from one dog to another by xonografting. Questionable ? 1908, filtered extracts from Chicken leukemia cells induced tumors in other birds

Figure 3.2 The Biology of Cancer (© Garland Science 2007) 1910, Peyton Rous discovered that the Rous Sarcoma virus (RSV) is able to induce tumors in chicken 1913, Dane Johannes Grib Figiber reported that stomach cancers in rats could be traced to spirochete worms that they harbored. Thus, he received Nobel Prize, direct validation of the idea that cancer was an infectious disease But Metastatic stomach epithelia in those rats were present in the normal stomach with Vitamin deficient. Misinterpretation? Fraud ? Things easily go wrong Since then, Peyton Rous report had been disregarded for two decades

Figure 3.6 The Biology of Cancer (© Garland Science 2007) In the Lab of Renato Dulbercco in CalTech, Harry Rubin found that stocks of RSV infected in Chicken Embryonic Fibroblast and some of these cells survived indefinitely. The RSV infected cells in cultures showed many traits of cancers. Form a foci, round shape, highly metabolic stage etc

Figure 3.6 The Biology of Cancer (© Garland Science 2007) In the Lab of Renato Dulbercco in CalTech, Graduate student Howard Temin showed RSV can transform normal cells to cancer cells. Cancer formation could be studied at the level of individual cells and these processes could be traced easily Loss of contact inhibition, foci formation Howard Temin showed the ability of RSV to transform cells in vitro.

Figure 3.7b The Biology of Cancer (© Garland Science 2007) Transformation In vitro Proposed the processes of transformation Loss of contact inhibition, foci formation, an anchorage independent growth Tumorigenicity can be tested in a host nude mice

Figure 3.8 The Biology of Cancer (© Garland Science 2007) Question is whether RSV is required for maintenance of transformation or simply initial infection is sufficient ? The continued presence of RSV is needed to maintain transformation How prove it ? Tem-sensitive mutant and the maintenance of transformation Viral transforming gene is required to both initiate and maintain the transformed phenotype of virus- infected cells ? This finding is also true In vivo ?

Figure 3.3 The Biology of Cancer (© Garland Science 2007) Viruses have simple life cycles

Figure 3.16 The Biology of Cancer (© Garland Science 2007) Retroviral genome become integrated into the chromosome of infected cells How did RSV RNA virus succeed in transmitting in genome through many generations within a cell lineage ? Late 1960, Temnin proposed a radical idea explaining that RSV made ds DNA from RNA and become a part of chromosomal DNA by integration How was DNA made from RNA ? Central Dogma can not explain this phenomenon ? Reverse trasnscriptase was found within a year Retrovirus re-infection can explain the stable transformation ? No. RSV virus with no ability to replicate was still able to transform cells

Figure 3.19 The Biology of Cancer (© Garland Science 2007) Src gene from RSV is responsible for transformation Retrovius has small number of genes (gag, poly,env genes. Several mutant of RSV –No replication with transformation activity –Replication without transformation activity In 1974, Bishop and Varmus defined the transformation-associated sequences of RSV genome by making probes. How did they found ? Src gene was found, but interestingly, this src was also clearly present in the uninfected chicken cells ? There are two copies of the src-related gene in normal cells C-src play a role in cellular processes in normal cells, but v-src is able to transform normal cells to tumors

Figure 3.22 The Biology of Cancer (© Garland Science 2007) ALV (avian leukosis virus) may kidnapp cellular Src gene during lysogenic life cycle and become RSV ? Viral Oncogene would be changed, acquire the tranformation activity What is the different action of Cellular oncogene and Viral Oncogene ? How one Viral Oncogene can induce multiple change during tumorigenesis ? C-oncogene might be modified in many non-viral induced tumors ? How do other retrovirus without oncogene induce tumors ?

Retroviruses cause cancer by kidnapping mutated versions of normal cellular genes

The structures of c-src and v-src and Deletion of the C-terminal region leads to activation of v-Src Binds peptides phosphorylated on Tyr Binds polyproline motifs

Figure 3.9a The Biology of Cancer (© Garland Science 2007) DNA viruses are able to induce cancer 1960, Shope found that papillomas can be induced a virus (papillomavirus). This Virus contain DNA molecule Several tumor DNA tumor viruses were found. Papillomavirus, the mouse polyomavirus and SV40 are grouped together as the Pavovavirus (papilloma, polyoma and Vacuoles); Circular ds DNA Adenovirus and herpesvirus have long linear ds DNA

Figure 3.10a The Biology of Cancer (© Garland Science 2007) SV40 tumor virus SV40 virus (the 40 th simian virus monkey virus) was discovered as a contaminant of the polyovirus vaccine stocks SV 40 induces cell lysis in permissive host cells, but can not replicate in Nonpermissive cells and instead transform these cells SV40 contaminant might trigger cancer (mesothelioma) in many people who were vaccinated, but no evidence ?

Table 3.2 The Biology of Cancer (© Garland Science 2007)

Figure 3.14 The Biology of Cancer (© Garland Science 2007) Tumor Virus become part of host cell DNA in virus transformed cells Discovery of tumor virus-associated proteins were found in cancers induced by DNA virus; indicating that the viral genome place in tumor cells encoded a protein (SV40 T Ag) How could viral genomes be replicated and transmitted for many generations of cells ?The virus are replicated as extranchromosomal molecules in permissive cells. 1968, found that the viral DNA in SV40- transforned cells was associated with chromosomal DNA, suggesting a integration of viral DNA into chromosome Viral genome become a part of cell’s genome. T Ag in cancer

Integrated SV40 genomes How can you find this integrated SV40 genome in chromosomes ?

Table 3.3 The Biology of Cancer (© Garland Science 2007)

Figure 3.23a The Biology of Cancer (© Garland Science 2007) How do other retrovirus without any oncogene induce tumors ? Slowly tranforming retrovirus activates proto-oncogene by inserting their genomes adjacent cellular genes Avian Leukosis virus(AVL) or murine leukemia virus (MVL) with no concogen induce a malignancy ? Map the site of integration of viral genome. Myc was a target site of viral integration of tumors induced by these viruses ? Insertinal mutagenesis Is it possible to transform cells with random integration of DNA or Viral DNA in vitro ? These insertional mutagenesis provided a powerful tool to find new oncogenes

Retroviruses also cause cancer by randomly inserting next to, thus activating the expression of proto-oncogenes exons Transcribe to mRNA 5 kilobases Retroviral insertion sites in different tumors

Table 3.4 The Biology of Cancer (© Garland Science 2007)

Some retroviruses naturally carry oncogene HTLV (human T cell leukemia virus) were transmitted via milk-borne from mother to infants. In Kysu, 1 % infection in the population. Tax gene product from viral genome appears to activate the two cellular genes (IL-2, GM-CSF), which stimulates the proliferation of hematopoietic cells Gene therapy can induce the tumors Immune deficiency (AIDS) can induce the some types of viral-induced tomors ‘ Kaposi’s Sarcoma in endotherial lining of lymphduct. Related with human herpesvirus-8 (HHV-8). How about the helicobacter ? HBV, HCV ?

Summary Retrovirus The causative agents of transplantable tumors in chickens, mice, and rats RNA virus The virus carries genetic information responsible for transforming a normal cell into tumor cell. Retrovirus : RNA-containing animal virus that replicate through a DNA intermediate : Two classes based on the latent period of tumorogenesis 1) Acute transforming retrovirus rapidly produce tumors in newborn animals and carry genetic information capable of inducing tumors directly. 2) The slowly transforming retroviruses do not carry oncogenes and induce tumors by integrating themselves adjacent to a cellular gene and altering its transcriptional regulation.