我的學思歷程 ～從冠狀病毒到肝炎病毒～ 賴明詔 Michael M.C. Lai 成功大學校長 April 17, 2009

成功大學 成立於 1931 於台南 （鄭成功 的故鄉 ) 工學院，理學院，文學院， 電機資訊，規劃設計， 管理學院，醫學院，社會科學，生物科技學院 學生 : 名 (10462 名大學生， 6948 名碩士， 3128 名博士 教授 : 1789 名 職員 : 3673 名 (1520 名於大學部， 2153 名於附設醫院 ) 台灣第二大大學 企業界最愛

我的研究生涯 大學時代 ( 台大 醫學院醫科 ) 研究生時代 ( 加大柏克萊 ) 大學教授時代 ( 南加大 ) 腫瘤病毒 冠狀病毒 肝炎病毒 (D and C 型） 回國 (2003) 後 中央研究院副院長 成功大學校長

病毒形狀

病毒形狀 ( 二 )

廿面體的基本構造 Triangulation numbers 5-, 3-, 2x symmetry Quasisymmetry

Structure of Calicivirus

病 毒 的 基 因病 毒 的 基 因 ☼ 人 類 細 胞 有 3 萬 個 基 因 ☼ 細 菌 有 5 千 個 基 因 ☼ 病 毒 有 10~100 個 基 因

The Central Dogma DNA RNA Proteins RNA

Retroviruses (RNA tumor viruses) (Viruses with reverse transcriptase) Rous Sarcoma virus (Peter Duesberg, Harry Rubin)  RNA, glycoproteins, phosphoproteins  Oncogenes: src.  Rel (NFkB), erb, etc. (Peter Vogt)

Demyelinating diseases Multiple sclerosis: central nervous system demyelination (virus-induced autoimmune reaction)  Theiler’s encephalomyelitis virus (picornavirus)  Measles virus  Experimental allergic encephalitis (EAE)  Mouse hepatitis virus (MHV): Coronavirus

Coronavirus （冠狀病毒 )

Multiple sclerosis 多發性神經硬化 Common colds 傷風感冒 SARS

Genome structure of Coronavirus: 31-kb, single-strand, (+)-sense RNA

HESEMN (A)n 31 kb POL MHV Genomic and Subgenomic RNAs ( Leader Intergenic Sequence 3'-UTR RNA recombination

台灣 SARS 病例

肝 炎 病 毒肝 炎 病 毒  A 型 : 急性肝炎 Picornavirus  B 型 : 慢行肝炎 ( 肝癌 ) Hepadnavirus  C 型 : 慢性肝炎 ( 肝癌 ) Flavivirus  D 型 : 慢性肝炎 Deltaviridae  E 型 : 急性肝炎 Calicivirus

Hepatitis Delta Virus RNA Nucleocapsid (HDAg) HBsAg Major protein Middle protein Large protein Phospholipid

Challenges of Hepatitis Delta Virus Viroid-like circular RNA with a protein Make both full-length RNA replication and subgenomic mRNA transcription Carry out RNA-dependent RNA transcription, but do not encode RNA-dependent polymerase:  Cellular RdRP?

Hepatitis C virus 170 million carriers in the world (2% of population) Tendency to cause persistent infection, leading to chronic hepatitis, liver cirrhosis, hepatoma A flavivirus with single-stranded RNA

Hepatitis C virus Relative risks of hepatocellular carcinoma in non-Asians of Los Angeles County HBV (-), HCV (-)1.0 HBV (+)5.1 HCV (+)22.3 HCV (+), HBV (+)56.2 (Adapted from Yuan et al. [1999])

HCV Genes and Gene products Lindenbach et al, Nature 933, 2005 ER

Life cycle of HCV Nature 436, (18 August 2005 ) ER or Golgi ? rER ? Endosome Receptor Golgi ?

HCV Genome and HCV Replicon R. Bartenschlager et al. / Antiviral Research 60 (2003) 91–102

Clinical manifestations of HCV Liver:Asymptomatic or acute hepatitis  chronic hepatitis  liver cirrhosis  hepatocellular carcinoma B cells:Mixed cryoglobulinemia (B-cell oligoclonal proliferation)  non-Hodgkin’s B-cell lymphoma Autoimmune diseases: Sjogren’s syndrome Pseudomembranous glomerulonephritis Purpura, Diabetes mellitus, thyroid disease

HCV HCV: A non-retroviral oncogenic RNA virus Frequent Mutations of p53 (Teramoto et al., Cancer Res.) B cell Lymphoma Hepatocellular Carcinoma Hypermutation of Ig Ivanovski et al., Blood) Chronic Infection Liver Cirrhosis Lymphoproliferative Diseases (mixed Cryoglobulinemia)

Establishment of a B-cell lymphoma cell line (SB cells) Established from the spleen of an HCV-positive patient with B-cell lymphoma and mixed cryoglobulinemia: proof that HCV infects B cells in vivo. Produces infectious virus particles that are capable of infecting B cells but not hepatocytes. The immunoglobulin gene is monoclonal but undergoes continuous evolution

Molecular mechanism of viral lympho- vs hepatotropism Harvest Detection (qRT-PCR) In vitro Transcribed RNA Electroporation into Huh7.5 and Raji

SB virus RNA, but not JFH-1 Virus, Replicates in Raji Cells   Raji

JFH-1 Virus, but not SB Virus RNA, Replicates in Huh7.5 Cells   Huh7.5 SB virus is lymphotropic. JFH-1 virus is hepatotropic.

HCV induces a mutator phenotype HCV infection causes a 5-10-fold increase in mutation frequency of cellular genes, including immunoglobulin (Ig) and somatic genes, e.g., p53 or  -catenin genes. Increased mutations are seen in in vitro HCV-infected B cells and in PBMC from HCV-infected individuals. Mutations are amplified in the HCV-infected lymphoma and hepatoma.

Amplified oncogene mutations in HCV(+) B-cell tumors and hepatoma B cell lymphomas Hepatocellular carcinomas (HCC) HCV-associated Non-HCV HCV tumor nontumor HBV tumor nontumor Nonviral tissues Mutation frequency (mutation/bp) x10 -4 p53  -catenin

MK Day p.i. Raji JT DSBs VHVH HCV RNA Mock HCV (+) HCV (-) DW MK - HCV+ - HCV+ D C B A E F DSBs in V H DSBs in p53 DSBs Double-stranded DNA breaks in HCV- infected cells (by linker-ligation PCR)

Challenges of Hepatitis C Virus Vaccines Therapy (interferon and ribavirin) Mechanism of HCV pathogenesis (persistent infection) and oncogenesis  The mechanism of immune escape  The role of B and T cell infection

Challenges of Infectious Diseases Antibiotics Small pox Polio Hepatitis B virus Human papillomavirus HIV Malaria Drug-resistant tuberculosis

Emerging and reemerging infectious diseases Courtesy of Dr. Anthony Fauci, NIAID SARS Avian flu HIV Marburg Measles

大學生要學什麼 追求自己的興趣，不管「熱」或「冷」門 不怕選「錯」門 多嘗試新的領域 重視通識教育，培養人文藝術素養及品德 養成表達的能力