© life_edu Lecture 31 Part IVb. Cancer Biology and Emergent Treatments: Applications of Biotechnology in Cancer Treatment Issues in Biotechnology: The

© life_edu

Lecture 31

Part IVb. Cancer Biology and Emergent Treatments: Applications of

Biotechnology in Cancer Treatment

Issues in Biotechnology:The Way We Work With Life

Dr. Albert P. Kausch

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Medical Biotechnology

Issues in Biotechnology:Biotechnology, Our Society and Our Future

OnCampus LiveOnCampus LiveBCH 190, MIC 190, AFS 190, NRS 190, PLS 190BCH 190, MIC 190, AFS 190, NRS 190, PLS 190

OnLine BCH 190OnLine BCH 190

A Sweeping General Survey on Life and BiotechnologyA Public Access College Course

The University of Rhode Island

Kimberly Nelson



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© life_edu

A Sweeping General Survey on Life and Biotechnology

The University of Rhode Island



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BCH 190BCH 190 Section II.

The Applications of Biotechnology

© life_edu

Lecture 31

Part IVb. Cancer Biology and Emergent Treatments: Applications of

Biotechnology in Cancer Treatment



life edu.us

Medical Biotechnology

Cancer Basics, and the Application of Biotechnology to Treat Disease

Emergent Technologies in Cancer Treatment

Cancer: Presentation Overview - Part 2

Fundamentals of Oncology • Socio-economic burden of cancer • Origins of cancer (carcinogenesis)• Pathophysiology and Molecular pathology• Contemporary target discovery/validation

Applications of Biotechnology in Cancer Treatment• A brief history of cancer chemotherapy• Three Case Studies - New Drugs Making a Difference• Issues and opportunities for doing even more

The complex interaction between a microbial pathogen and a host is the underlying basis of The complex interaction between a microbial pathogen and a host is the underlying basis of infectious disease. By understanding the molecular details of this interaction, we can identify infectious disease. By understanding the molecular details of this interaction, we can identify virulence-associated microbial genes and host-defense strategies and characterize the cues to virulence-associated microbial genes and host-defense strategies and characterize the cues to which they respond and mechanisms by which they are regulated. which they respond and mechanisms by which they are regulated.

Microarrays consist of ordered Microarrays consist of ordered sets of DNA fixed to solid sets of DNA fixed to solid surfaces provide pharmaceutical surfaces provide pharmaceutical firms with a means to identify firms with a means to identify drug targets. drug targets.

In the future, the emerging In the future, the emerging technology promises to help technology promises to help physicians decide the most physicians decide the most effective drug treatments for effective drug treatments for individual patients. individual patients.

DNA Microarray A technology that is reshaping molecular biology

This information will guide the design of a new generation of medical tools.This information will guide the design of a new generation of medical tools.

How Does Chip Technology Work?How Does Chip Technology Work?

RNA expression

Nature 415: 530-536 2002

Disease free

Metastases

• Genetic causes of interpatient variability in phenotype

• Relationship between patient phenotype and genotype

Phenotype: Clinical symptomsPharmacokinetic variabilityResponse to drug

- efficacy- side effects

Genotype: A genetic marker(s) distinguishing specific variations within a DNA

sequence

Non-respondersResponders

Pharmacogenomics

Human Genetics

• SNPs• Haplotypes• Sequencing

Expression Profiling

• Specific transcript levels• Total RNA profiling

Proteomics

• Specific biochemical markers

• Protein profiling

Phenotype• Drug response

• Disease

Prediction

Pharmacogenomics

Whan that Aprill, with her shoures sooteThe droghte of March is perced to the rooteAnd bathed every veyne in swich licour,Of which vertu engendred is the flour;Whan Zephirus eek with his sweete breethInspired hath in every holt and heethThe tendre croppes, and the yonge sonneHath in the Ram his halfe cours yronne,And smale foweles maken melodye,And slepen al the nyght with open eye-(So priketh hem Nature in hir corages);Thanne longen folk to goon on pilgrimagesAnd especially from every shires endeto room 10, in CBLS they wende,

Whan that Aprill, with her shoures sooteThe droghte of March is perced to the rooteAnd bathed every veyne in swich licour,Of which vertu engendred is the flour;Whan Zephirus eek with his sweete breethInspired hath in every holt and heethThe tendre croppes, and the yonge sonneHath in the Ram his halfe cours yronne,And smale foweles maken melodye,And slepen al the nyght with open eye-(So priketh hem Nature in hir corages);Thanne longen folk to goon on pilgrimagesAnd especially from every shires endeto room 10, in CBLS they wende,

When in April the sweet showers fallThat pierce March’s drought to the root And bathed every vein in sweet liquor That has power to generate therein and sire the flower;When Zephyr also has with his sweet breathFilled again, in every holt and heath,The tender shoots and leaves, and the young sunHis half-course in the sign of the Ram has run,And many little birds make melodyThat sleep through all the night with open eye(So Nature pricks them on to ramp and rage)Then folk do long to go on pilgrimagesAnd especially from every shire’s end Of URI they to 271 Chaffee they went

Variation in PhenotypeVariation in Phenotypeisis

Variation in GeneticsVariation in GeneticsXX

Variation in Variation in EnvironmentEnvironment

Vp = Vg X Ve

Complex (Multifactorial) Phenotypes: What can we expect?

Genei

Genen

Environment

I. Few Genes each contributing large risk• ls> 4.0

II. Many genes each contributing small risk• ls< 2.0

Gi

E

Gi

GiGi

Gi

Expression ofPhenotype =Gi + Gn + E

Phenotype Phenotype

Genetics Genetics X X

EnvironmentEnvironment

What Causes Cancer?What Causes Cancer?

Genetics Genetics

EnvironmentEnvironment

Mutations Oncogenes Anueploidy Chromosomal Rearrangements


Genetics Genetics

Human Cancer Genes: Genes Causally Linked to Oncogenesis

About 290 genes have been linked to cancer by identification of mutation in primary human tumors. Changes include…

Point mutations Activating (ex. Ras in pancreatic cancer) Inactivating (ex. p53 in ~50% of all cancers)

Translocations Aberrant fusion proteins (ex. bcr/abl in CML)High Expression (ex. erbB2 in breast cancer)

Deletions/insertions/frame shifts (ex. APC in colon cancer)

Point mutationsFrame shift mutationsDeletionsInsertionTranslocationsAdditional gene copiesAnueploidyChromosomal rearrangementsOncogene activationsViruses (e.g. HPV)

Step-wise (ordered) Genetic Changes in Colon Tumorigenesis

“Full Blown” Tumorigenesis requires 4-17 events (mutations)

Kinzler and Vogelstein Cell 1996

EnvironmentEnvironmentCarcinogens (Too many to name)Toxic compounds (everywhere)

Sunlight (UVb)Tobacco smoking

Radiation


Emergent Technologies in Cancer Treatment: Lecture Overview


Emergent Technologies in Cancer TreatmentApplications of Biotechnology in Cancer Treatment

• A brief history of cancer chemotherapy• Three Case Studies - New Drugs Making a Difference• Issues and opportunities for doing even more

Emergent Technologies in Cancer Treatment: Lecture Overview



• USS John Harvey, an American ship in Bari Harbor, carried a highly classified load of 2,000 100-lb mustard bombs• Dec 2, 1943 - A German raid damaged 17 ships, including the Harvey• Fire on the Harvey caused a mustard-laden smoke that spread quickly• 617 Mustard gas poisoning cases among troops and merchant marine seamen, with a 14% fatality rate (3-fold higher that of World War I)• Commonly cited as the key observation that led to Goodman and Gilman’s pioneering work on the use of nitrogen mustards in cancer chemotherapy

The Bari Myth

R. Joy, HISTORICAL ASPECTS OF MEDICAL DEFENSE AGAINST CHEMICAL WARFARE

After World War I, researchers were highly aware of the human effects of mustard gas—it destroyed lymphatic tissue and bone marrow. It was broadly reasoned that mustards could also kill cancer cells in the lymph nodes.

It was not until 1942, more than a year before the Bari raid, however, that two young assistant professors in Yale’s new Department of Pharmacology, Louis S. Goodman, M.D., and Alfred Gilman, Ph.D., were commissioned by the Defense Dept. to study nitrogen mustard and its derivatives. Their pre-clinical studies in mice produced dramatic regressions of lymphomas.

In December of 1942, a 48-year-old patient with terminal lymphosarcoma was given 10 consecutive doses of nitrogen mustard, a 10th of a milligram to a milligram per kilogram of body weight, roughly 2.5 times what became the standard dose.

Within two days a softening of the tumor masses was noted. By the end of treatment the tumors disappeared. A month later, however, the patient relapsed, and subsequent courses of treatment were less effective. Nevertheless the scientists were encouraged.

The clinical trials remained a classified military secret, even from caregivers, until 1946—the first Yale patients’ charts said only, “0.1 mg. per kg. compound X given intravenously.” R

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74:

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( 200

1 )The Bari Myth (cont’d)

R Ruddon, Anticancer Drugs

From Humble Beginnings…

Goodman and Gilman usher in 60 years of cytotoxic chemotherapy

R Ruddon, Anticancer Drugs

1st Line 2nd Line

Breast Ca Adriamycin+ Cytoxan +/- Taxol

Cytoxan + 5FU+methotrexate

Cytoxan + Adriamycin + 5FU

Taxol + Herceptin (erbB2+ only)

Tamoxifen

Taxol

Xeloda

Herceptin

Taxotere

NSCLC Taxol and carboplatin

Gemcitabine + CDDP

Tarceva, Taxotere

Etoposide, Navelbine

Colorectal Camptosar + 5FU+ leucovorin

Oxaliplatin + 5FU+leucovorin

Xeloda

Irinotecan

Prostate LHRH analog (Lupron/Zoladex)

+/- antiandrogen

Casodex+mitoxantrone+pred-nisone

Hodgkins Lymphoma ABVD: doxorubicin+bleomycin+ vinblastine+ dacarbazine

Cytotoxic Drugs in 2012 - Still Among Standards of Care in Cancer Chemotherapy

Tumor Type Doubling time (days)

Burkitt's lymphoma 2 - 5 daysTesticular cancer 21Ewings' sarcoma 22Small cell lung cancer 81Colon cancer 96Breast cancer 129Lung cancer 134

The key limitation of cytotoxic chemotherapy…

How fast is the tumor growing?

Populations of committed stem cells in normal tissues(bone marrow; epithelium of GI tract) have doubling times < 1 day

Cancer: Presentation Overview



Inhibition of Angiogenesis as a Target

Required for tumor metastasis & growth beyond 1-2 mm3

Limited role in adults Broad spectrum of

activity May circumvent

acquired drug resistance

Folkman: Angiogenesis is Necessary

VEGFVascular Epithelial Growth Factor

Angiogenesis and Tumors

Blood vessels grow toward and into tumors

DNA TechnologyDNA TechnologyAnd Small Molecule And Small Molecule Drug DesignDrug Design

Precise treatments for:

Cancers Cardiovascular diseases Inflammation

Behavior Obesity Depression Schizophrenia

Small Molecule Cancer Drug Design

Small Molecule Cancer Drug Design

Gleevec: Imatinib, is a drug used to treat certain cancers. It is marketed by Novartis as Gleevec (USA) or Glivec (Europe/Australia/Latin America) as its mesylate salt, imatinib mesilate (INN).

Imatinib is the first of a new class of drugs that act by specifically inhibiting a certain enzyme – a receptor tyrosine kinase – that is characteristic of a particular cancer cell, rather than non-specifically inhibiting and killing all rapidly dividing cells.

Recombinant DNA And Recombinant DNA And New Cancer Drugs New Cancer Drugs

DNA TechnologyDNA Technologyand Pharmaceuticalsand Pharmaceuticals

Allows precise treatments for:

Cancers Cardiovascular diseases Inflammation

Behavior Obesity Depression Schizophrenia

Promoter Coding Sequence Terminator

Genetic Constructs Now Make Proteins That Are New Cancer Drugs

Controlled expression“making protein”

Your favorite gene

Stop transcriptionMessage stabilityErbituxTM

HerceptinTM

AvastinTM

RituxanTM

Advance 1: AvastinTM (Bevisizumab)

Why an Antibody to VEGF?

Ferrara: VEGF is Necessary for Angiogenesis

Ideal for extracellular targets Small molecule approaches successful for “beautiful” binding sites, but

generally limited for all others– Non-enzymes– Non-receptors– Shallow pocket enzymes

Opportunity for exquisite selectivity– Enzymatic selectivity for targets with promiscuous binding sites

IGF-1R & Insulin receptor – Eliminates potential for chemotype-based toxicities associated with small

molecules Relatively long duration of coverage over target

– T 1/2 for human antibodies (weeks vs. hours for drugs)

Antibodies for targeted therapyAdvantages

Distribution of Abs may limit activity relative to small molecules– Distribution: potential for limited tumor penetration

Single chain variable fragments may improve penetration– Potential for immunogenicity

Fully human antibodies may mitigate risk Manufacturing more complicated; high COG Long T1/2: Little opportunity to limit exposure if toxic

– Points to advantage for small molecule, or Ab fragments Human Abs rarely x-react with rodents: preclinical modeling is

challenging– Efficacy modeling for anti-angiogenesis targets– Estimates of TI in the efficacy species

Antibodies for targeted therapyPotential Issues/Risks

Therapeutic Antibodies for Cancer Antibodies already a key weapon in the Cancer arsenal Regulatory approvals for Abs in solid tumor indications are on the rise

Product Target Company Indication

Approval date

RituxanTM CD20 IDEC/Roche/

Genentech

NHL

11/97

HerceptinTM erbB2 Genentech/

Roche

Metastatic BC

9/98

AvastinTM VEGF Genentech 1st line CRC

2/2004

ErbituxTM EGFR ImClone/BMS 2nd line CRC

2/2004

Breast Cancer:Phase III study of Avastin plus paclitaxel in first-line metastatic breast

doubled the duration of surviving without cancer progression compared to chemotherapy alone

Median progression-free survival was 11 months for patients treated with Avastin plus chemotherapy, compared to six months for patients treated with chemotherapy alone

In patients with measurable disease, the overall response rate was 28 percent (93/330) in the Avastin plus chemotherapy arm, vs. 14

percent (45/316) observed in the chemotherapy alone armKD Miller, ASCO 2005

Lung Cancer:Phase III study of Avastin plus paclitaxel and carboplatin chemotherapies

in first-line non-small cell lung cancer (NSCLC) patients showed a 30 percent improvement in overall survival (12.5 months vs. 10.2

months)compared to patients who received chemotherapy aloneAB Sandler, ASCO 2005

Additional Benefit from Avastin

Advance 2: HerceptinTM (traztuzumab)

v-erbB v-erbA5’ 3’

AEV

Heregulins

EGFTGF-

AmphiregulinBetacellulin

HB-EGF

NRG2NRG3

HeregulinsBetacellulin

ErbB-1 (EGFR) ErbB-2 ErbB-3 ErbB-4

Extracellular Ligand-binding Domain

Tyrosine Kinase Domain

• erbB2 (HER2) is a member of the epidermal growth factor receptor (EGFR) family of transmembrane tyrosine kinases • Amplification (an excess number of gene copies) or over-expression

(excess production of protein) confers on the affected cancer cell aggressive behavioral traits, including enhanced growth and proliferation, increased invasive and metastatic capability, and stimulation of angiogenesis

• Patients with breast cancer in which HER2 is amplified (FISH-positive tumors) or HER2 is over-expressed are likely to have poorly differentiated tumors with a high proliferative rate, positive axillary lymph nodes, and decreased expression of estrogen and progesterone receptors

erbB2 as a Cancer Target

NE J Med 353: 1734 (2005)Gabriel N. Hortobagyi, M.D.

erbB2 as a Target: ChecklistHerceptinTM (traztuzumab)

1. Is the target a ‘gain-of-function’ gene that provides a key advantage to the tumor:• Progression and expression are linked?• Inhibition of gene function restores normal

homeostasis?2. Is the target gene differentially expressed in tumor

vs. normal cells (plausible basis for a TI?)3. Can the target be measured in biopsy material to

select patients for clinical treatment?4. Can modulation of the target after Rx be monitored

by a relatively non-invasive technique (biomarker)?5. Does target structure/function indicate a plausible

basis for therapeutic attach (Is it an enzyme? A receptor?)

6. Is it plausible that inhibition of the target would combine with other known Rx?

/ ?

/ ?

Herceptin (anti-erbB2)Monoclonal Antibody: Biological Effects

ErbB-2

LA Emens, Amer J Therapeutics 12: 243 (2005)

Herceptin: Clinical Activity in Advanced Breast Cancer

LA Emens, Amer J Therapeutics 12: 243 (2005)

Herceptin – Activity in Improving Survival(D + C, TX =/- trastuzumab)

Romond et al., NE J Med 353: 1673 (2005)

“On the basis of these results, our care of patients with HER2-positive breast cancer must change today. Certainly, patients with lymph-node–positive,

HER2-positive breast cancer should receive trastuzumab as part of optimal adjuvant systemic therapy… Since most HER2-positive tumors have other adverse prognostic factors, this risk–benefit scenario is likely to apply to many patients with node-negative breast cancer.”

NE J Med 353: 1734 (2005)Gabriel N. Hortobagyi, M.D.

Issues in BiotechnologyIssues in Biotechnology

One of the standards of care in cancer treatments include the use of chemotherapy. Many of the drugs used in modern chemotherapy are:

(A) carcinogenic (B) cytotoxic (C) not developed by large pharmaceutical companies (D) free of any side-effects (E) deadly

Therapeutic Antibodies for Cancer Antibodies already a key weapon in the Cancer arsenal Regulatory approvals for Abs in solid tumor indications are on the rise

Product Target Company Indication

Approval date

RituxanTM CD20 IDEC/Roche/

Genentech

NHL

11/97

HerceptinTM erbB2 Genentech/

Roche

Metastatic BC

9/98

AvastinTM VEGF Genentech 1st line CRC

2/2004

ErbituxTM EGFR ImClone/BMS 2nd line CRC

2/2004

Issues in BiotechnologyIssues in Biotechnology

Antibodies have some distinct advantages over old school chemotherapies for treating cancer because they:

(A) all of these reasons (B) are ideal for extracellular targets involved with tumor growth (C) eliminate potential for chemotype-based toxicities associated with small molecules (D) have a relatively long duration of coverage over the tumor target

EGFR(erbB1)

Advance 3: TarcevaTM (erlotinib)

Why Target EGFR for Anti-tumor Drug Discovery ?

• Over-expression of EGFR can transform cells in a ligand-

dependent manner

• EGFR or TGFa are frequently over-expressed in carcinomas, for example 42% of Lung Cancers over-express EGFR and many express EGF or TGFa

• EGFR “Knockout” Mice are viable and hematopoietic cells lack EGFR

The Use of Mouse AsThe Use of Mouse As A Human Cancer Model A Human Cancer Model

The Use of Mouse Genomics AsThe Use of Mouse Genomics As A Human Cancer Model A Human Cancer Model

OncoMouse

OncoMouse

Pharmacological Selectivity - Among close oncogene kindreds

N

N

HN

ON

NH

O

N

N

HN

O

N

N

HN

O

O

N

N

HN

ON

O

O

OO

CP-358774erbB2 kinase: 1714 nM

EGFR kinase: 2 nM857x selective for EGFR

erbB2 kinase: 43 nMEGFR kinase: 18 nM

erbB2 kinase: 40 nMEGFR kinase: 5998 nM

CP-724714erbB2 kinase: 8 nM

EGFR kinase: 6173 nM772x selective for erbB2

OMe

represents >500000xshift in selectivity!

HN

Extracellular targets Small molecules successful for

“beautiful” binding sites Opportunity for exquisite

selectivity Relatively long duration of

coverage over target– T 1/2 for human antibodies

(weeks vs. hours for drugs)

Antibodies Advantages Tarceva Intracellular targets Tyrosine Kinases active

sites Exquisite selectivity

(500,000-fold vs. erbB2) Relatively long duration

of coverage over target– T 1/2 in humans ~12

hours… q.d. oral dosing (strong patient preference)

High tumor penetration Low COG

New Frontiers - Tumor Stem Cells

BCRP/ABCG2 (breast cancer resistance protein)

Telomerase

Oct4

Nanog

Stat3

Aldehyde Dehydrogenase

CXCR4/SDF-1

• Targeted therapies, those abrogating the key molecular advantages of tumor cells, are adding importantly to patient survival

• These approaches offer the promise of less morbidity and higher quality of life

• Monoclonal antibodies have ‘credentialed’ targeted therapies; small molecule drugs are now emerging as well

• The future is likely to be focused on understanding ‘systems biology’ vs. individual gene targets, on tumor stem cells vs. their diverse (and somewhat less talented) progeny, and on novel combinations of targeted agents where patient survival will be measured in decades, not months

Cancer: Summary for Part 2

No Walls

The Clear bead at the center changes everythingThere are no edges to my being now

I have heard it said that there is a windowThat opens from one mind to another

But where there are no wallsThere is no need for a window, or fitting a latch.

Rumi 1279 AD

For those who are interested in taking this For those who are interested in taking this course for college credit through the course for college credit through the

University of Rhode Island; University of Rhode Island; For more information please contact:For more information please contact:

[email protected]@gmail.com

CreditsCredits

Lectures by: Lectures by:

Edited by:Edited by:

Video Produced by:Video Produced by:

Thank You to The University of Rhode Island Thank You to The University of Rhode Island and all of the students of Issues in and all of the students of Issues in

Biotechnology over the yearsBiotechnology over the years

Dr. Albert KauschDr. Albert Kausch

Dr. Albert Kausch Dr. Albert Kausch and Kimberly Nelsonand Kimberly Nelson

Thaddeus WeaverThaddeus Weaver

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© life_edu Lecture 31 Part IVb. Cancer Biology and Emergent Treatments: Applications of Biotechnology in Cancer Treatment Issues in Biotechnology: The