1. Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658 Henrietta Lacks Cancer: Detection and Treatment

2. Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658 Henrietta Lacks: the source of HeLa cells

3. Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658 Henrietta Lacks: the source of HeLa cells “During the 1970s and 1980s, as many as one in three cell lines deposited in cell culture repositories were imposters, one cell line overtaking or masquerading as another. The most notorious culprit was a cervical carcinoma line, HeLa, established by George Gey at the Johns Hopkins Medical School in 1951”

4. Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658 Henrietta Lacks: the source of HeLa cells An estimated $10 million of research was discredited.

5. Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658 Henrietta Lacks: the source of HeLa cells DNA fingerprinting has been used to differentiate different cell lines. “cell lines from the same person showed 0.3–2.9% fragment differences”

6. HeLa cell variation J. Cell Biol. Bottomley et al. 41 (3): 806

7. Fig. 3 TRENDS in Ecology and Evolution Vol 21 pg 47 Balance between Longevity and Health

8. Mutations

9. Cancer: is the loss of control over cell division. Tumors are normal cells that are dividing inappropriately. –They stop performing their “normal” function, and are dividing repeatedly.

10. A cell becomes cancerous when there are incorrect positive AND negative signals.

11. GO! STOP! cancer

12. Multiple mutations are required for cancer to occur Fig 22.17

13. Tbl 22.10

14. Causes of mutations: Replication errors –Exacerbated by poor DNA repair –Limited by telomere length Other biological agents –Viruses –Transposons Environmental factors –Ultraviolet light –Mutagenic chemicals smoking, industrial waste, natural toxins

15. Environment plays a large role in the chance of contracting cancer… The multiethnic cohort study: exploring genes, lifestyle and cancer risk. L Kolonel, D Altshuler, B Henderson (July 2004) Nature Reviews Cancer 4, 519-527 Fig 1

16. The rapidity of increased childhood thyroid cancer in the heavily contaminated areas of Belarus, Ukraine and Russia was surprising. 4 years

17. Source: US Mortality Public Use Data Tape 2000, National Center for Health Statistics, Centers for Disease Control and Prevention, 2002. 1.Heart Diseases710,76029.6 2.Cancer553,09123.0 3.Cerebrovascular diseases167,661 7.0 4.Chronic lower respiratory diseases122,009 5.1 5.Accidents (Unintentional injuries) 97,900 4.1 6.Diabetes mellitus 69,301 2.9 7.Influenza and Pneumonia 65,313 2.7 8.Alzheimer’s disease 49,558 2.1 9.Nephritis 37,251 1.5 10.Septicemia 31,224 1.3 RankCause of Death # of deaths % of all deaths US Mortality, 2000

18. Change in the US Death Rates* by Cause, 1950 & 2000 * Age-adjusted to the 2000 US standard population. Source: US Mortality Volume 1950, National Vital Statistics Report, 2002, Vol. 50, No. 15. Heart Diseases Cerebrovascular Diseases Pneumonia/ Influenza Cancer 1950 2000 Rate Per 100,000

19. 2003 Estimated US Cancer Cases* *Excludes basal and squamous cell skin cancers and in situ carcinomas except urinary bladder. Source: American Cancer Society, 2003. Men 675,300 Women 658,800 32%Breast 12%Lung & bronchus 11%Colon & rectum 6%Uterine corpus 4%Ovary 4%Non-Hodgkin lymphoma 3%Melanoma of skin 3%Thyroid 2%Pancreas 2%Urinary bladder 20%All Other Sites Prostate33% Lung & bronchus14% Colon & rectum11% Urinary bladder6% Melanoma of skin4% Non-Hodgkin lymphoma4% Kidney3% Oral Cavity3% Leukemia3% Pancreas2% All Other Sites17%

20. 2003 Estimated US Cancer Deaths* ONS=Other nervous system. *Excludes basal and squamous cell skin cancers and in situ carcinomas except urinary bladder. Source: American Cancer Society, 2003. Men 285,900 Women 270,600 25%Lung & bronchus 15%Breast 11%Colon & rectum 6%Pancreas 5%Ovary 4%Non-Hodgkin lymphoma 4%Leukemia 3%Uterine corpus 2%Brain/ONS 2%Multiple myeloma 23%All other sites Lung & bronchus31% Prostate 10% Colon & rectum 10% Pancreas5% Non-Hodgkin4% lymphoma Leukemia4% Esophagus4% Liver/intrahepatic3% bile duct Urinary bladder3% Kidney3% All other sites22%

21. Cancer Death Rates*, for Men, US, 1930-1999 *Age-adjusted to the 2000 US standard population. Source: US Mortality Public Use Data Tapes 1960-1999, US Mortality Volumes 1930-1959, National Center for Health Statistics, Centers for Disease Control and Prevention, 2002. Lung Colon and rectum Prostate Pancreas Stomach Liver Rate Per 100,000 Leukemia

22. Cancer Death Rates*, for Women, US, 1930-1999 *Age-adjusted to the 2000 US standard population. Source: US Mortality Public Use Data Tapes 1960-1999, US Mortality Volumes 1930-1959, National Center for Health Statistics, Centers for Disease Control and Prevention, 2002. Lung Colon and rectum Uterus Stomach Breast Ovary Pancreas Rate Per 100,000

24. Tobacco Use in the US, 1900-1999 *Age-adjusted to 2000 US standard population. Source: Death rates: US Mortality Public Use Tapes, 1960-1999, US Mortality Volumes, 1930-1959, National Center for Health Statistics, Centers for Disease Control and Prevention, 2001. Cigarette consumption: Us Department of Agriculture, 1900-1999. Per capita cigarette consumption Male lung cancer death rate Female lung cancer death rate

26. Tobacco Use in the US, 1900-1999 *Age-adjusted to 2000 US standard population. Source: Death rates: US Mortality Public Use Tapes, 1960-1999, US Mortality Volumes, 1930-1959, National Center for Health Statistics, Centers for Disease Control and Prevention, 2001. Cigarette consumption: Us Department of Agriculture, 1900-1999. Per capita cigarette consumption Male lung cancer death rate Female lung cancer death rate

29. Treating cancer: Avoid it –Avoid mutagens –DNA repair gets less efficient as we age

30. T-cells recognize and eliminate abnormal cells; such as cells with many mutations Our immune system protects us from cancer

31. P53 is activated by DNA damage Fig 22.15

32. p53 can induce apoptosis via two pathways: Nuclear and/or Mitochondrial

33. Treating cancer: Avoid it –Avoid mutagens –DNA repair gets less efficient as we age Surgery –Must remove all cancer cells –Non-invasive

34. Treating cancer: Avoid it –Avoid mutagens –DNA repair gets less efficient as we age Surgery –Must remove all cancer cells –Non-invasive Radiation –Directed at tumor; causes DNA damage -> cellular self-destruction –Mutagenic, side effects

35. Treating cancer: Avoid it –Avoid mutagens –DNA repair gets less efficient as we age Surgery –Must remove all cancer cells –Non-invasive Radiation –Directed at tumor –Mutagenic, side effects Chemotherapy –Toxins directed at rapidly dividing cells –Mutagenic, many side effects

36. Chemotherapy a rapidly dividing cell Toxin XX

37. Normal Multi-Drug Resistance protein MDR toxin/hormone/etc

38. Some cancers over-express MDR Toxin MDR toxin I’m a cancer cell with over-expressing MDR. I laugh at your toxins.

39. The Epigenetic Progenitor Origin of Human Cancer (2007) A P Feinberg, R Ohlsson, S Henikoff Nature Reviews Genetics 7: 21-31 Mutations continue after cancer develops

40. O O OOO O O OO OO Cancer cell with mutation causing MDR over-production Evolution: changes in DNA as information transmitted

41. O O OOO O O OO OO O O OOO O O OO OO Apply chemo- therapy X XX XXXX X XX Kills most cells. Except if some have mutation that allow them to be resistant. Evolution: changes in DNA as information transmitted Cancer cell with mutation causing MDR over-production

42. O O OOO O O OO OO O O OOO O O OO OO O X XX XXXX X XX Kills most cells. Except if some have mutation that allow them to be resistant. Continues to replicate Evolution: changes in DNA as information transmitted Apply chemo- therapy Cancer cell with mutation causing MDR over-production

43. O O OOO O O OO OO O O OOO O O OO OO O O OOO O O OO OO O X XX XXXX X XX Kills most cells. Except if some have mutation that allow them to be resistant. Continues to replicate Tumor with cells expressing MDR Evolution: changes in DNA as information transmitted Apply chemo- therapy Cancer cell with mutation causing MDR over-production

44. Some cancers over-express MDR Toxin MDR toxin I’m a cancer cell with over-expressing MDR. I laugh at your toxins.

45. Cells need the proximity of blood vessels to survive One of the latest chemotherapy treatments involves cutting off the blood supply to the tumor. (anti-angiogenesis)

46. Tumors must have sufficient blood flow to continue cell division induce blood vessel growth (angiogenesis)

47. Tumors Evolve: Only tumor cells near blood vessels or that can attract blood vessels survive.

48. Targeting toxins to cancer cells… A vesicle with mutant genes that cause blood vessels to die is directed to newly growing blood vessels by interacting with Integrins. Integrins are present on newly growing blood vessels, but not on established blood vessels.

49. Detecting Cancer or Types of Cancer

50. Cancer Cells Normal Cells

51. A Microarray is a chip with DNA sequences (genes) bound to the surface at known locations. It can be used to track or monitor expression of many genes.

52. Tracking changes in gene expression using a Microarray

53. Making cDNA from RNA

54. Tracking changes in gene expression using a Microarray

56. Use of microarray to estimate genes likely present in malignant cancers

57. Patients cancer free for 5+ years Patients cancer spread in 5 years Use of microarray to estimate genes likely present in malignant cancers similar to Fig 22.19 different genes

58. Fig 22.19 Microarrays can be used to get information about types of cancers

59. Young (>55) Breast cancer patients More accurate profiling of tumors results in more accurate choices of treatments. Patients with benign tumors can avoid chemotherapy (adjuvant).

60. Effect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari, K M Lonergan, R T Ng, C MacAulay, W L Lam, and S Lam BMC Genomics, 8:297

61. Effect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297 CS=current smoker, FS=former smoker, NS=never smoked Table 1: Subject Demographics

62. Overlapping and unique genes expression Fig 1B Effect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297

63. Table 3: Reversible gene expression upon smoking cessation related to mucus secretion (genes in bold have not been previously associated with smoking)

64. CABYRENTPD8TFF3 Fig 4A Effect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297 Some changes in gene expression induced by smoking are reversible

65. MUC5ACGSK3B Fig 4B Effect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297 Smoking can induce irreversible changes in gene expression

66. Treating cancer: Avoid it –Avoid mutagens –DNA repair gets less efficient as we age Surgery –Must remove all cancer cells –Non-invasive Radiation –Directed at tumor –Mutagenic, side effects Chemotherapy –Toxins directed at rapidly dividing cells –Mutagenic, many side effects

67. Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658 Henrietta Lacks Cancer: Detection and Treatment