Sydney Cancer Institute

1
Sydney Cancer Institute

• University of Sydney 19th Jan 2004

2
Organisation and Research Direction

• The Sydney Cancer Institute is an independent
  research institute established in February 2003
  by
• Sydney Cancer Centre
• The Central Sydney Area Heath Service (CSAHS)
• The University of Sydney
• The Sydney National Cancer Foundation
• The primary function of the institute is
• Conducting research to substantially improve
  cancer outcomes

3
Objectives and Research Directions

• The Sydney Cancer Institutes research programs
  are based around a clinically driven set of
  research priorities
• The Sydney Cancer Institute comprises the
  following divisions
• Molecular Oncology
• Cancer Biology
• Experimental Therapeutics
• Cancer Care and Control

4
Research Goals

• The Sydney Cancer Institute aims to pursue
  research excellence, depth and relevance by
• Integration with clinical cancer medicine by the
  development of translational research programs
• Providing core research facilities such as
• A clinical cancer register
• Statistician and information technology
• Shared instruments and operators
• A tumour bank available to all SCI researchers
• Providing more structured national and
  international connections

5
Scientific Advisory Board

• The SCI scientific advisory board includes
  researchers with a diverse array of skills and
  interests
• Prof. W McCarthy (Director Melanoma Foundation)
• Prof. J Bishop (Director Sydney Cancer Centre)
• Prof. B Armstrong (Head School of Public Health,
  USyd)
• Prof. G Halliday (Department of Dermatology,
  USyd)
• Prof. P Lay (Centre for Heavy Metals Research,
  USyd)
• Prof. B Roufogalis (Pro-Dean Pharmaceutical
  Chemistry, USyd)
• Prof. J Simes (Director NHMRC Clinical Trials
  Centre, USyd)
• Prof. J Thompson (Director Sydney Melanoma Unit)
• A/Prof. P Buttow (Executive Director, Medical
  Psychology Unit, USyd)
• A/Prof. M Millward (Head Clinical Research, SCI)
• Dr S Clarke (Senior Research Fellow, SCI)

6
Affiliations

• The SCI has affiliation agreements with
• The University of Sydney
• Central Sydney Area Health Service
• The Sydney National Cancer Foundation
• The John Wayne Cancer Institute Los Angeles
• The National Institutes of Health (USA)
• National Cancer Institute (CTEP, USA)
• Cancer Therapeutics Research Group (Singapore)
• National Cancer Centre (Singapore)
• John Hopkins (Singapore)

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Research programs

• The SCI has a diverse research program including
• DNA Methylation (Dr Susan Clarke)
• Cancer Genetics and Drug Resistance (Dr Quihan
  Dong, Dr John Young)
• Viral Oncology (Professor Y Cossart and Dr Carol
  Thompson)
• Tumour Pathology and Molecular Biology (A/Prof.
  Soon Lee)
• Skin Cancer Biology (Prof. Gary Halliday)
• Skin Cancer and Photobiology (Dr Vivienne Reeve)
• Cancer Invasion and Metastasis (Dr Guy Lyons)
• Heavy Metals Research Program (Prof. Peter Lay)
• Clinical Pharmacology (A/Prof. Stephen Clarke)
• Pharmaco Oncology (Prof. Basil Roufogalis)
• Gene Therapy (A/Prof John Rasko)
• Molecular Imaging (A/Prof Michael Fulham)
• Clinical Trials (A/Prof. Michael Millwood)
• Early Detection and Diagnosis (Dr Scott Menzies)

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Biometals Section

• Centre for Heavy Metals Research
• School of Chemistry
• University of Sydney

9
Biometals Section, CHMR

• Associate Professor Robert Armstrong
• Dr. Rachel Codd
• Dr Carolyn Dillon
• Dr. Ron Fenton
• Professor Hans Freeman
• Professor Trevor Hambley
• Associate Professor Margaret Harding
• Dr. Hugh Harris
• Associate Professor Brendan Kennedy
• Professor Peter Lay
• Professor Len Lindoy
• Associate Professor Tony Masters
• Dr. Lou Rendina

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Research Areas/Interests

• Anti-Cancer Drugs
• - Boron complexes boron neutron capture
  therapy (Rendina)
• - Co(III) Hypoxic Agents (Hambley)
• - Copper Complexes (Dillon, Hambley, Harris,
  Kennedy, Lay)
• - Metallocenes (Dillon, Harding)
• - Metal Complexes of Organic Anti-Cancer Drugs
  (Hambley,
• Harding)
• - Metalloporphyrin/Fullerenes (Armstrong, Lay)
• - Pt(IV) and Pt(II) (Fenton, Hambley)
• - Ruthenium (Armstrong, Dillon, Lay)
• Radiopharmaceuticals
• - Use of copper-64 in macrocyclic systems for
  the imaging
• and therapy of cancer (Lindoy)
• - Improved 99Tc Generators (Masters)

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Research Areas/Interests

• Metal-Induced Cancers and Toxicity
• - Cr- and Ni-induced cancers (Codd, Dillon,
  Harris, Lay)
• - As carcinogenesis and toxicity (Dillon,
  Harris)
• Anti-Inflammatory Drugs
• - Cu, Zn, Ni, Zn complexes as
  anti-inflammatories (Dillon,
• Hambley, Kennedy, Lay)
• Anti-Diabetics
• - Cr(III) (Dillon, Harris, Lay)
• - Vanadium (Codd, Lay)

12
Research Areas/Interests

• Metalloproteins
• - structure of heme proteins and their roles in
  the immune system, and heart disease (Armstrong,
  Lay)
• Chelation Therapy and Metabolic Processes
• - role of transition metal-sialic acid species
  in metal
• homeostasis/disease (Codd)
• - cold-adapted 'super-siderophores' for metal
  chelation
• therapies (Codd).
• Diagnostics
• - use of vibrational spectroscopy in the
  diagnosis of cancer
• (Armstrong, Lay)

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Facilities/Expertise

• Structural Biology and Structural Chemistry
• X-ray, Neutron Electron Scattering and
  Diffraction Synchrotron Techniques NMR
  Spectroscopy See separate presentation on CSBSC
• Mass Spectrometry
• Electrospray (including HPLC front end) GC-MS
  Maldi
• Vibrational Spectroscopy
• IR, Raman and Resonance Raman including tissue
  mapping
• EPR Spectroscopy
• L, Q and X band, ENDOR, He cryogenics, Whole cell
• Cell Biology
• Cytotoxicity Genotoxicity Permeability
  Imaging, Spectroscopy
• Animal Studies
• Pharmacology Pharmacokinetics Efficacy
  Toxicity

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Anti-Cancer Drugs
15
Boron Neutron Capture - Rendina
2
0

energy
(2.28 MeV)
5
5
3
16
Clinical BNCT Agents
Malignant brain tumours Malignant melanoma

• Two new classes
• Platinum(II)-amine complexes
• Metallo-intercalator complexes

17
Dinuclear Platinum Complexes
Woodhouse, S. L. Rendina, L. M. Chem. Commun.,
2001, 2464. International Patent PCT/AU02/00943
18
Metallo-intercalator Complexes
Todd, J. A. Rendina, L. M. Inorg. Chem., 2002,
41, 3331. International Patent PCT/AU02/00943.
19
Control A375 Melanoma Cells
J. Aitken, H. Harris, P. A. Lay, USyd, P. Farmer,
UC Irvine
20
CuDSF-treated A375 Melanoma Cells
J. Aitken, H. Harris, P. A. Lay, USyd, P. Farmer,
UC Irvine
21
Cu XANES of Melanoma Cells
J. Aitken, H. Harris, P. A. Lay, USyd, P. Farmer,
UC Irvine
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Selective activation in solid tumours Hambley
necrotic
Pt(IV)
suboxic
normoxic
23
MicroXANES of Pt obtained from A2780 ovarian
cancer cells treated with Pt(IV)
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Identification of Novel Intercalating Platinum
Compounds - Fenton, Aldrich-Wright

• CISPLATIN analogs bind to DNANew Pt compounds
  intercalate into DNA - IP protection- PCT
  filing, Priority date February 22nd 2001

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EFFICACY of lead compound - Comparison with
Cisplatin in Cisplatin-resistant cell lines

L1210 mouse leukemia 2008 human ovarian
tumor PC3 prostate tumor DDP
Cisplatin-resistant C13 acquired SKOV3
intrinsic Cisplatin resistance

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Titanocene Dichloride
Phase I clinical trials toxic effects
hypoglycaemia, metallic after taste bone
marrow largely unaffected liver toxicity
does-limiting side-effect
Phase II clinical trials breast cancer
renal cell carcinoma

• poor hydrolytic stability in water pHgt 4.0
• Ti accumulates in nucleic rich regions tumour
  cells
• species formed in vivo and how interaction with
  DNA occurs not understood
• mechanism is distinct from platinum based drugs

A/Prof Margaret Harding, School of Chemistry
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Antitumour Metallocenes
Ti
V
Mn
Cr
Tc
Zr
Nb
Mo
Re
Hf
Ta
W
Y Cl, Br, I, NCS, N3

• Each complex has independent mechanism of action
• Current studies focus on cellular distribution,
  interaction with biomolecules of Mo, Nb
  complexes
• M Mo targets thiols

Design water soluble, stable derivs
A/Prof Margaret Harding, School of Chemistry
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Streptonigrin

• Clinical use human cancers until 1977
• broad spectrum antitumor activity
• lymphoma, melanoma
• cancers of the breast, cervix, head, neck
• severe and unpredictable side-effects

Water, pH 6.5
A/Prof Margaret Harding, School of Chemistry
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Streptonigrin Metal Complexes
affects DNA binding topo II recognition site
Redox related to DNA cleavage ACCELERATED
BY METAL IONS
bipyridyl
zwitterion
labile metal complexes

• Ru(II) complex
• Reductively activated to semiquinone
• Strong DNA binding/cleavage predicted

A/Prof Margaret Harding, School of Chemistry
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Radiopharmaceuticals
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Improved Tc-99 Generators - Masters

• Understanding the interactions between alumina
  and molybdates has led to improvements in the
  process for generating 99Tc for radio-diagnostics
  

Schematic outline of 99Mo production process at
ANSTO.
32
Metal-Induced Cancers and Toxicity
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Cellular Metabolism of Chromium
34
Effect of Oxidation State on Cr Genotoxicity
Cr(VI)
Cr(V)
Cr(III)
Dillon, C. T. et al. Chem. Res. Toxicol. 1998,
11, 119-129 Dillon, C. T. et al. Chem. Res.
Toxicol. 2000, 13, 742-748.
35
Exposure of Whole Cells to Cr(VI) and Cr(III)
Cr(III)-Treated Cell
P
K
Cr
Zn
Cr(VI)-Treated Cell
P
K
Cr
Zn
Min
Max
Dillon, C. T. Lay, P. A. Kennedy, B. J.
Stampfl, A. P. J. Cai, Z. Ilinski, P.
Rodrigues, W. Legnini, D. G. Lai, B. Maser, J.
J. Biol. Inorg. Chem. 2002, 7, 640-645.
36
Anti-Inflammatory Drugs
37
Copper Indomethacin Lay, Hambley, Kennedy, Dillon

• Copper indomethacin is a dimeric copper complex
  containing 4 indomethacin ligands bound to Cu
  through the carboxylic acid group.
• Cu-algesic (CuIndo) is an effective
• anti-inflammatory drug commonly used in dogs and
  horses.
• Importantly, CuIndo is much less TOXIC in dogs
  than IndoH.

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Assessment of Small Intestinal Ulceration
39
Effect of the Formulation on Gastro-Intestinal
Damage

• Equivalent doses of indomethacin (10 mg/kg) were
  administered to each animal.
• The number of animals tested per treatment ranged
  from 4-6.

40
Anti-Diabetics

• Codd, Harris, Lay

41
Efficacy of Cr Dietary Supplements

• Chromium supplements are the second biggest
  market for dietary supplements (over 1B
  industry).
• Taken to convert fat into muscle in athletes
  (used instead of steroids for humans) and food
  animals.
• Used to help prevent diabetes.
• There is no compelling evidence (epidemiology,
  cell work, or biochemical assays) that Cr
  supplements convert fat into muscle or help
  prevent diabetes.
• The FDA has prevented companies from advertising
  such health benefits.
• There are anti-diabetic effects exhibited in
  animal studies for certain Cr complexes and they
  are believed to have anti-diabetic effects on
  humans.

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Side-Effects of Cr Dietary Supplements

• They react with enzyme systems such as glucose
  oxidase and xanthine oxidase to form highly
  genotoxic mixtures of high oxidation state Cr
  species, which are very damaging to DNA
• The Cr(VI) generated in these enzymatic processes
  and strongly inhibits phosphatase enzymes, which
  is likely to be responsible for the anti-diabetic
  effects of Cr supplements
• High-oxidation-state Cr and V species appear to
  act in the same way, through similar
  intermediates in their anti-diabetic effects

Inhibition of PTP (phosphatase enzyme by Cr(VI),
Cr(V) and V(V). Mulyani, Levina, Lay, JACS,
submitted
43
Metalloproteins
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Heme Proteins - Armstrong, Harris, Lay

• Characterized NO, CO and O2 adducts of heme
  proteins, many of which are too unstable to
  crystallise
• Studied unfolding of cytochrome c
• Collaborations with Paul Witting and Roland
  Stocker
• - Studies on indolamine 2,3-dioxygenase (IDO) -
  important in the immune system
• - Studies on the roles of heme proteins in heart
  disease

45
Heme Proteins
Rich, Cheng, Armstrong and Lay
46
Chelation Therapy and Metabolic Processes
47
Metal-Sialic Acid Speciation Codd
Insight into the nature and role of species
formed between sialic acid and transition metal
ions. This polyfunctional carbohydrate is
present as the terminal residue in many
glycopoteins involved in metal transport (e.g.,
transferrin ceruloplasmin)
Membrane glycoproteins and glyco- lipids
extracellular sugar residues on mammalian plasma
membranes
48
Metal-Sialic Speciation
Extension of studies of metal-sialic acid
speciation with other biologically relevant
transition metal ions (e.g., Cu, Fe, Zn, V).
Cu(II)-sialic acid profile
Cr(V)-sialic acid profile
Implications for metal transport, homeostasis,
and role(s) in sialylglycoprotein-dependent
disease
Development of transition metal EPR Spectroscopy
as a diagnostic technique for glycosylation
patterns
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Diagnostics
50
Vibrational Spectroscopy in Breast Cancer
Diagnosis - Tam, Armstrong, Carter, Lay
51
Current Diagnostic TechniquesTriple Assessment
Side Effects

• A combination of physical examination, mammogram
  and fine needle aspiration cytology.
• Standard for breast diagnosis.
• Side effects
• Probable false positive and negative results from
  mammogram
• Time consuming progress

52
New Diagnostic For Breast Cancer

• 1H Magnetic Resonance Spectroscopy at IMRR
• Simple and quick diagnosis of aspirated breast
  tissues
• The choline-to-creatine peak ratio is being used
  to compare various breast disease states
• Vibrational Spectroscopy
• Infrared Spectroscopy
• Quick diagnosis by comparing the spectral
  differences in proteins (amides), lipids and DNA
  levels of sectioned tissues
• Raman Spectroscopy
• By using various laser excitations to compare
  proteins, lipids and DNA levels of different
  breast diseases.
• Imaging and mapping provide parallel results
  often difficult to determine in pathology.

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1H Magnetic Resonance Spectroscopy
ppm
54
FT-IR Spectra
Malignant
Transmission
Benign
Wavenumbers (cm-1)
55
FT-IR Spectra
Transmission
Malignant
Wavenumbers (cm-1)
56
Malignant
Raman Intensity (a.u.)
Benign
FT-Raman Spectra
Wavenumber (cm-1)
57

• Raman Imaging
• New approach to compare results parallel to the
  native pathological examinations
• Colour intensity of particular Raman marker
  bands have potential for identifying the progress
  of cancer
• Proteins, lipids, DNA and other important
  components can be detected in tissues that
  require less preparation than for infrared and
  magnetic resonance spectroscopy

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Summary

• 1H Magnetic Resonance Spectroscopy
• Fast and suitable for low-lipid content of breast
  lesions
• Research is in collaboration with The Institute
  for Magnetic Resonance Research, University of
  Sydney
• Vibrational Spectroscopy
• Suitable for both high- and low-lipid content of
  breast lesions
• Inexpensive and non-destructive.
• can run in parallel with histopathology
• Multivariate Statistical Analysis
• Improve significance for the spectral data
• Correlate and classify different breast diseases