The Iron Metabolism and Chelation Program - CCIA
description
Transcript of The Iron Metabolism and Chelation Program - CCIA
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The Iron Metabolism and Chelation Program - CCIA
Des Richardson
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Aims of the Iron Metabolism and Chelation Program(1) To develop new therapeutic agents to treat cancer and a variety of other diseases.
(2) To understand how cancer and normal cells metabolise iron this knowledge is critical for achieving goal #1
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Techniques UsedWide Range of Techniques Used: Chemistry and Biology
Synthetic and Inorganic Chemistry - Collaboration with P. Bernhardt (U.Q.)
X-ray crystallography and molecular modeling
Cell Biology
Molecular Biology Simple to Complex (Gene Knockout Studies in Mice to Determine Molecular Function )
Animal Studies- Analysing Effects of Compounds in Vivo in Animal Models
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Projects Underway in the Iron Program
1. Development of novel anti-metabolites: iron chelators/metal complexes with selective anti-tumour activity
2. Examining the role of iron in the cell cycle p21 and p53
3. Examination of the function of the melanoma tumour antigen, melanotransferrin- generation of melanotransferrin KO mouse.
4. Examination of the cytotoxic effects of anthracyclines via their avid interaction with cellular iron
5. The effects of nitric oxide and carbon monoxide on Fe metabolism. NO has a high affinity for Fe and plays a role in the anti-tumour effects of macrophages (a role for CO ??)
6. Development of iron chelators for the treatment of Friedreichs Ataxia and -Thalassaemia and understanding the function of frataxin in intracellular Fe trafficking.
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Development of New ChelatorsDesign of novel chelators is required for commercial interest and clinical application
Provisional Patent Hybrid Iron Chelators
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New NT series analoguesNTN2mTN4mTN44mTN4eTN4aTN4pTSTThiosemicarbazones - NT seriesHydrazones311311m - NNHN44pHNoctH effect of increasingly lipophilic substituents at terminal N4
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Some PIH analogues markedly inhibit tumor cell growthRichardson D, Tran E, Ponka P Blood (1995)
Pyridoxal - 100 seriesSalicylaldehyde - 200 series2-Hydroxy-1-naphthaldehyde - 300 series
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Selective anti-proliferative activity
efflux
3.50693183770.1820638981
7.41471903890.6185408672
43.64671204811.0110996768
41.27784325060.9172270926
3.17903572130.2462487945
12.93220284590.6142202956
4.41681565450.3103442422
28.98766324721.3863013841
43.26727938140.9459199834
34.85951462782.0327302508
36.06188774611.8365922503
37.26110391180.9013484931
27.70191009140.1538082955
35.69730951351.2223406338
Fe uptake 22.2.00
% Cellular Iron Released
effluc TC
2.731846317510.744623312964.969932998719.74343674864.41064018251.06669882491.06669882490.74804382540.74804382540.27096307170.27096307171.879076851.879076850.68985204990.6898520499
3.839762840421.83607994571.830945954931.510971413869.99795298161.45610905611.45610905611.59483454661.59483454660.83750398930.83750398930.84334338580.84334338580.27517510430.2751751043
6.962486854441.371010879378.468884731245.940835788276.43375906721.23248493921.23248493921.19167495611.19167495610.36256068640.36256068640.34061423560.34061423560.4365638650.436563865
10.649926782654.875444758180.378032606753.907218528481.03815488653.73662454633.73662454631.28040970681.28040970680.64698383140.64698383141.06219697981.06219697980.99268792080.9926879208
Control
DFO
311
NT
N44mT
Time (h)
% Cellular Iron Released
uptake
1004.2
92.62165206144.28
6.66700876320.94
9.46381226840.56
95.88901370359
90.14222784212.8818577655
90.98.1
41.953.3712474269
4.970.288267196
19.642.6852675951
18.742.0933098294
25.352.1496329757
25.358.5636126411
38.82372683451.2023389055
Internalized Iron Uptake (% control)
uptake TC
10090.50275466648.280279882989.04327844575.44336658426.28289349796.282893497913.608381911413.60838191145.15827503555.15827503550.30598498460.30598498461.25697177491.2569717749
10084.74642645835.36161963982.3944966243.78245486985.66824721975.66824721976.92268937756.92268937753.21668697663.21668697660.59273445570.59273445570.86497459860.8649745986
10075.91294755854.13544646375.11757666542.69832989465.95443972935.95443972932.58650513082.58650513089.03465428319.03465428310.12984510570.12984510570.29040154550.2904015455
10047.63132696811.631420494560.62028416260.67914372042.30621680382.30621680381.44351145051.44351145051.86579133091.86579133090.0554884710.0554884710.48459841780.4845984178
Control
DFO
311
NT
N44mT
Time (h)
Internalized Iron Uptake (% control)
MTT
0.1101.9404639287110.862125264497.2354058355104.9141939268128.920653707898.370699989292.25754223394.317077289494.8043811677100.6973960731112.896045037497.6901608582101.7433031097
0.293.693476925772.002881808590.138965084989.486296821396.745117676582.651088282997.63388544990.439568596990.97111.30670876694.3869100455109.072633638698.7695575086
0.3940.434940399633.705492360392.5655.891796777697.327809896755.5859507208102.855019961384.295716274584.113738050798.412571076281.5103.7673831976102.6235184192
102.038605285522.4613.773181107478.2631.963311159986.812036236226.499995097871.358426093773.682039329984.780730623990.225328154793.7112.957045914693.5352921497
92.226196435413.13688471857.928909812762.579814247216.674.976100514413.409640079948.363349937651.508125656967.946427408193.256533121696.188644201987.832807529996.7665844068
96.410.63993131436.161.68.481.13078708977.696607573318.404687351328.33085988739.044121359250.921654156896.681.696716567184.6423594758
88.69.96.653.6780892639786.16561205448.17100853679.620264427222.838642790626.887150449211.458283012175.316085860529.643362877466.9312856893
56.67.84115572925.308965429351.55152782225.475779897764.03241225478.02972009288.68409473517.40481483217.891502760210.155699067640.44939064599.958504008961.8961895217
DFO
311
NNH
ST
NT
N2mT
N4mT
N44mT
N4eT
N4aT
N4pT
NOctH
N44pH
FeNT
Chelator Concentration (mM)
Cellular Proliferation (% control)
MTTMRC51
0.1101.9404639287104.9141939268128.920653707898.370699989292.2575422330.192.6174496644100.0583089522107.125243570694.403198175896.7415732506
0.293.693476925789.486296821396.745117676582.651088282997.6338854490.289.115844762297.1966868704108.8498564152103.283268587994.0952103656
0.3940.434940399655.891796777697.327809896755.5859507208102.85501996130.3995.8856142399101.5539348621107.10403499196.251645117898.3389849717
102.038605285522.4631.963311159986.812036236226.499995097871.358426093790.235714148183.7467172454104.9878563539103.7669430193101.6428110784106.2921032183
92.226196435413.136884718516.674.976100514413.409640079948.363349937691.198054774690.2830463963107.6426712987110.3259988661100.0240211224103.0557669922
96.410.63993131438.481.13078708977.696607573318.404687351386.289956934766.2386927342106.4242803477111.8752133616100.763901805389.3963823523
88.69.9786.16561205448.17100853679.620264427283.039164584660.1692442369101.8314546437105.6021913722105.932619046185.6532706677
56.67.841155729212.564.03241225478.02972009288.68409473580.679212909775.576305806898.8238078623111.649294796497.695573033371.0267890763
72.363922414675.576305806885.0602317064111.306727347692.081543233569.8859743024
MRC-5Fibroblasts
SK-N-MCNeuroepithelioma
DFO (S)
311 (S)
NT (S)
N2mT (S)
N4mT (S)
N44mT (S)
DFO (F)
311 (F)
NT (F)
N2mT (F)
N4mT (F)
N44mT (F)
Chelator Concentration (mM)
Cellular Proliferation (% control)
NTpapMRC52
0.1101.9404639287110.8621252644104.9141939268128.920653707898.370699989292.2575422330.192.617449664493.048745313100.0583089522107.125243570694.403198175896.7415732506
0.293.693476925772.002881808589.486296821396.745117676582.651088282997.6338854490.289.115844762286.126334006397.1966868704108.8498564152103.283268587994.0952103656
0.3940.434940399633.705492360355.891796777697.327809896755.5859507208102.85501996130.3995.885614239993.5102394001101.5539348621107.10403499196.251645117898.3389849717
102.038605285522.4613.773181107431.963311159986.812036236226.499995097871.358426093790.235714148183.746717245488.2030573983104.9878563539103.7669430193101.6428110784106.2921032183
92.226196435413.13688471857.928909812716.674.976100514413.409640079948.363349937691.198054774690.283046396381.7421401788107.6426712987110.3259988661100.0240211224103.0557669922
96.410.63993131436.18.481.13078708977.696607573318.404687351386.289956934766.238692734270.6662820883106.4242803477111.8752133616100.763901805389.3963823523
88.69.96.6786.16561205448.17100853679.620264427283.039164584660.169244236969.743293914101.8314546437105.6021913722105.932619046185.6532706677
56.67.84115572925.308965429312.564.03241225478.02972009288.68409473580.679212909775.576305806873.665993654598.8238078623111.649294796497.695573033371.0267890763
72.363922414675.576305806865.8785.0602317064111.306727347692.081543233569.8859743024
MRC-5Fibroblasts
SK-N-MCNeuroepithelioma
DFO (S)
311 (S)
311m (S)
NT (S)
N2mT (S)
N4mT (S)
N44mT (S)
DFO (F)
311 (F)
311m (F)
NT (F)
N2mT (F)
N4mT (F)
N44mT (F)
Chelator Concentration (mM)
Cellular Proliferation (% control)
thymidine
101.135.533.792.589.152.392.3104.198.8126.687.812378.683.4
1414028.371.1109.372.279.747.6108.2100.352.5138.39386.2
156.140.537.285.570.976.4113.538.183.589.852.852.6106106.2
97.713.715.9111.964.676.449.2217.542.959.141.546.296.998.2
69.26574400254.3768819453.756468381660.596057363630.761789735954.084582610913.94010688192.412154061520.500415930924.64916970637.037853881527.8998701254.109184387590.7221177789
46.49504890713.4335932544.094484083885.15281598958.800784515837.39161791829.7933637532.5146677915.890397312321.22021843792.37744950264.699986616943.676176374112.5967714413
DFO
311
NNH
ST
NT
N2mT
N4mT
N44mT
N4eT
N4aT
N4pT
N44pH
NOctH
FeNT
Chelator Concentration (mM)
3H-Thymidine Incorporation (% control)
logPvsIC50
0.5
0.5
1.5
1.5
2.3
3.3
5.2
0.3
0.3
10.5
logP
log P calc
Chelator Antiproliferative Activity(IC50-mM)
mtttable
SK-N-MC neuroepitheliomaMRC-5 fibroblasts
DFO22>25
3110.3>25
311m0.3*
ST>12.5*
NT0.5>25
N2mT>12.5>25
N4mT0.5>25
N44mT1.5>25
N4eT1.6*
N4aT2.3*
N4pT3.3*
N44pH5.2*
NoctH10.5*
FeNT>12.5*
* not determined
logPtable
3112.17
311m2.17
ST0.64
NT1.76
N2mT2.72
N4mT2.19
N44mT2.3
N4eT2.4
N4aT2.61
N4pT3.75
N44pH*
NoctH5.19
* Unable to be calculated by Broto's method
Thytable
DFO23
3110.6
311m0.6
ST>25
NT9.0
N2mT15.5
N4mT6.3
N44mT1.5
N4eT5.7
N4aT7.9
N4pT3.8
N44pH4.3
NoctH17.4
FeNT>25
mttforsem
101.9404639287110.8621252644104.914193926898.370699989292.25754223394.317077289494.8043811677100.697396073197.690160858297.2354058355128.9206537078112.8960450374101.74330310970.1
93.693476925772.002881808589.486296821382.651088282997.63388544990.439568596990.97111.306708766109.072633638690.138965084996.745117676594.386910045598.76955750860.2
40.434940399633.705492360355.891796777655.5859507208102.855019961384.295716274584.113738050798.4125710762103.767383197692.5697.327809896781.5102.62351841920.39
22.4613.773181107431.963311159926.499995097871.358426093773.682039329984.780730623990.2253281547112.957045914678.2686.812036236293.793.5352921497102.0386052855
13.13688471857.928909812716.613.409640079948.363349937651.508125656967.946427408193.256533121687.832807529962.579814247274.976100514496.188644201996.766584406892.2261964354
10.63993131436.18.47.696607573318.404687351328.33085988739.044121359250.921654156881.696716567161.681.130787089796.684.642359475896.4
9.96.678.17100853679.620264427222.838642790626.887150449211.458283012129.643362877453.678089263986.165612054475.316085860566.931285689388.6
7.84115572925.30896542935.47577989778.02972009288.68409473517.40481483217.891502760210.15569906769.958504008951.551527822264.032412254740.449390645961.896189521756.6
311
311m
NT
N4mT
N44mT
N4eT
N4aT
N4pT
N44pH
ST
N2mT
NOctH
FeNT
DFO
Chelator Concentration (mM)
Cellular Proliferation (% control)
mtttabsem
SK-N-MC neuroepithelioma
3110.3
311m0.3
NT0.5
N4mT0.5
N44mT1.5
N4eT1.6
N4aT2.3
N4pT3.3
N44pH5.2
NoctH10.5
ST>12.5
N2mT>12.5
FeNT>12.5
DFO22
mtttablesem
SK-N-MC neuroepitheliomaMRC-5 fibroblasts
3110.3>25
311m0.3>25
NT0.5>25
N4mT0.5>25
N44mT1.5>25
N2mT>12.5>25
DFO22>25
FeNT>12.5*
* not determined
NTpapMRC5sem
101.9404639287110.8621252644104.914193926898.370699989292.257542233128.92065370780.192.617449664493.048745313100.058308952294.403198175896.7415732506107.12524357060.1
93.693476925772.002881808589.486296821382.651088282997.63388544996.74511767650.289.115844762286.126334006397.1966868704103.283268587994.0952103656108.84985641520.2
40.434940399633.705492360355.891796777655.5859507208102.855019961397.32780989670.3995.885614239993.5102394001101.553934862196.251645117898.3389849717107.1040349910.39
22.4613.773181107431.963311159926.499995097871.358426093786.8120362362102.038605285583.746717245488.2030573983104.9878563539101.6428110784106.2921032183103.766943019390.2357141481
13.13688471857.928909812716.613.409640079948.363349937674.976100514492.226196435490.283046396381.7421401788107.6426712987100.0240211224103.0557669922110.325998866191.1980547746
10.63993131436.18.47.696607573318.404687351381.130787089796.466.238692734270.6662820883106.4242803477100.763901805389.3963823523111.875213361686.2899569347
9.96.678.17100853679.620264427286.165612054488.660.169244236969.743293914101.8314546437105.932619046185.6532706677105.602191372283.0391645846
7.84115572925.308965429312.58.02972009288.68409473564.032412254756.675.576305806873.665993654598.823807862397.695573033371.0267890763111.649294796480.6792129097
75.576305806865.8785.060231706492.081543233569.8859743024111.306727347672.3639224146
MRC-5Fibroblasts
SK-N-MCNeuroepithelioma
311 (S)
311m (S)
NT (S)
N4mT (S)
N44mT (S)
N2mT (S)
DFO (S)
311 (F)
311m (F)
NT (F)
N4mT (F)
N44mT (F)
N2mT (F)
DFO (F)
Chelator Concentration (mM)
Cellular Proliferation (% control)
efflux
3.50693183770.1820638981
7.41471903890.6185408672
43.64671204811.0110996768
41.27784325060.9172270926
3.17903572130.2462487945
12.93220284590.6142202956
4.41681565450.3103442422
28.98766324721.3863013841
43.26727938140.9459199834
34.85951462782.0327302508
36.06188774611.8365922503
37.26110391180.9013484931
27.70191009140.1538082955
35.69730951351.2223406338
Fe uptake 22.2.00
% Cellular Iron Released
effluc TC
2.731846317510.744623312964.969932998719.74343674864.41064018251.06669882491.06669882490.74804382540.74804382540.27096307170.27096307171.879076851.879076850.68985204990.6898520499
3.839762840421.83607994571.830945954931.510971413869.99795298161.45610905611.45610905611.59483454661.59483454660.83750398930.83750398930.84334338580.84334338580.27517510430.2751751043
6.962486854441.371010879378.468884731245.940835788276.43375906721.23248493921.23248493921.19167495611.19167495610.36256068640.36256068640.34061423560.34061423560.4365638650.436563865
10.649926782654.875444758180.378032606753.907218528481.03815488653.73662454633.73662454631.28040970681.28040970680.64698383140.64698383141.06219697981.06219697980.99268792080.9926879208
Control
DFO
311
NT
N44mT
Time (h)
% Cellular Iron Released
uptake
1004.2
92.62165206144.28
6.66700876320.94
9.46381226840.56
95.88901370359
90.14222784212.8818577655
90.98.1
41.953.3712474269
4.970.288267196
19.642.6852675951
18.742.0933098294
25.352.1496329757
25.358.5636126411
38.82372683451.2023389055
Internalized Iron Uptake (% control)
uptake TC
10090.50275466648.280279882989.04327844575.44336658426.28289349796.282893497913.608381911413.60838191145.15827503555.15827503550.30598498460.30598498461.25697177491.2569717749
10084.74642645835.36161963982.3944966243.78245486985.66824721975.66824721976.92268937756.92268937753.21668697663.21668697660.59273445570.59273445570.86497459860.8649745986
10075.91294755854.13544646375.11757666542.69832989465.95443972935.95443972932.58650513082.58650513089.03465428319.03465428310.12984510570.12984510570.29040154550.2904015455
10047.63132696811.631420494560.62028416260.67914372042.30621680382.30621680381.44351145051.44351145051.86579133091.86579133090.0554884710.0554884710.48459841780.4845984178
Control
DFO
311
NT
N44mT
Time (h)
Internalized Iron Uptake (% control)
MTT
0.1101.9404639287110.862125264497.2354058355104.9141939268128.920653707898.370699989292.25754223394.317077289494.8043811677100.6973960731112.896045037497.6901608582101.7433031097
0.293.693476925772.002881808590.138965084989.486296821396.745117676582.651088282997.63388544990.439568596990.97111.30670876694.3869100455109.072633638698.7695575086
0.3940.434940399633.705492360392.5655.891796777697.327809896755.5859507208102.855019961384.295716274584.113738050798.412571076281.5103.7673831976102.6235184192
102.038605285522.4613.773181107478.2631.963311159986.812036236226.499995097871.358426093773.682039329984.780730623990.225328154793.7112.957045914693.5352921497
92.226196435413.13688471857.928909812762.579814247216.674.976100514413.409640079948.363349937651.508125656967.946427408193.256533121696.188644201987.832807529996.7665844068
96.410.63993131436.161.68.481.13078708977.696607573318.404687351328.33085988739.044121359250.921654156896.681.696716567184.6423594758
88.69.96.653.6780892639786.16561205448.17100853679.620264427222.838642790626.887150449211.458283012175.316085860529.643362877466.9312856893
56.67.84115572925.308965429351.55152782225.475779897764.03241225478.02972009288.68409473517.40481483217.891502760210.155699067640.44939064599.958504008961.8961895217
DFO
311
NNH
ST
NT
N2mT
N4mT
N44mT
N4eT
N4aT
N4pT
NOctH
N44pH
FeNT
Chelator Concentration (mM)
Cellular Proliferation (% control)
MTTMRC51
0.1101.9404639287104.9141939268128.920653707898.370699989292.2575422330.192.6174496644100.0583089522107.125243570694.403198175896.7415732506
0.293.693476925789.486296821396.745117676582.651088282997.6338854490.289.115844762297.1966868704108.8498564152103.283268587994.0952103656
0.3940.434940399655.891796777697.327809896755.5859507208102.85501996130.3995.8856142399101.5539348621107.10403499196.251645117898.3389849717
102.038605285522.4631.963311159986.812036236226.499995097871.358426093790.235714148183.7467172454104.9878563539103.7669430193101.6428110784106.2921032183
92.226196435413.136884718516.674.976100514413.409640079948.363349937691.198054774690.2830463963107.6426712987110.3259988661100.0240211224103.0557669922
96.410.63993131438.481.13078708977.696607573318.404687351386.289956934766.2386927342106.4242803477111.8752133616100.763901805389.3963823523
88.69.9786.16561205448.17100853679.620264427283.039164584660.1692442369101.8314546437105.6021913722105.932619046185.6532706677
56.67.841155729212.564.03241225478.02972009288.68409473580.679212909775.576305806898.8238078623111.649294796497.695573033371.0267890763
72.363922414675.576305806885.0602317064111.306727347692.081543233569.8859743024
MRC-5Fibroblasts
SK-N-MCNeuroepithelioma
DFO (S)
311 (S)
NT (S)
N2mT (S)
N4mT (S)
N44mT (S)
DFO (F)
311 (F)
NT (F)
N2mT (F)
N4mT (F)
N44mT (F)
Chelator Concentration (mM)
Cellular Proliferation (% control)
NTpapMRC52
0.1101.9404639287110.8621252644104.9141939268128.920653707898.370699989292.2575422330.192.617449664493.048745313100.0583089522107.125243570694.403198175896.7415732506
0.293.693476925772.002881808589.486296821396.745117676582.651088282997.6338854490.289.115844762286.126334006397.1966868704108.8498564152103.283268587994.0952103656
0.3940.434940399633.705492360355.891796777697.327809896755.5859507208102.85501996130.3995.885614239993.5102394001101.5539348621107.10403499196.251645117898.3389849717
102.038605285522.4613.773181107431.963311159986.812036236226.499995097871.358426093790.235714148183.746717245488.2030573983104.9878563539103.7669430193101.6428110784106.2921032183
92.226196435413.13688471857.928909812716.674.976100514413.409640079948.363349937691.198054774690.283046396381.7421401788107.6426712987110.3259988661100.0240211224103.0557669922
96.410.63993131436.18.481.13078708977.696607573318.404687351386.289956934766.238692734270.6662820883106.4242803477111.8752133616100.763901805389.3963823523
88.69.96.6786.16561205448.17100853679.620264427283.039164584660.169244236969.743293914101.8314546437105.6021913722105.932619046185.6532706677
56.67.84115572925.308965429312.564.03241225478.02972009288.68409473580.679212909775.576305806873.665993654598.8238078623111.649294796497.695573033371.0267890763
72.363922414675.576305806865.8785.0602317064111.306727347692.081543233569.8859743024
MRC-5Fibroblasts
SK-N-MCNeuroepithelioma
DFO (S)
311 (S)
311m (S)
NT (S)
N2mT (S)
N4mT (S)
N44mT (S)
DFO (F)
311 (F)
311m (F)
NT (F)
N2mT (F)
N4mT (F)
N44mT (F)
Chelator Concentration (mM)
Cellular Proliferation (% control)
thymidine
101.135.533.792.589.152.392.3104.198.8126.687.812378.683.4
1414028.371.1109.372.279.747.6108.2100.352.5138.39386.2
156.140.537.285.570.976.4113.538.183.589.852.852.6106106.2
97.713.715.9111.964.676.449.2217.542.959.141.546.296.998.2
69.26574400254.3768819453.756468381660.596057363630.761789735954.084582610913.94010688192.412154061520.500415930924.64916970637.037853881527.8998701254.109184387590.7221177789
46.49504890713.4335932544.094484083885.15281598958.800784515837.39161791829.7933637532.5146677915.890397312321.22021843792.37744950264.699986616943.676176374112.5967714413
DFO
311
NNH
ST
NT
N2mT
N4mT
N44mT
N4eT
N4aT
N4pT
N44pH
NOctH
FeNT
Chelator Concentration (mM)
3H-Thymidine Incorporation (% control)
logPvsIC50
0.5
0.5
1.5
1.5
2.3
3.3
5.2
0.3
0.3
10.5
logP
log P calc
Chelator Antiproliferative Activity(IC50-mM)
mtttable
SK-N-MC neuroepitheliomaMRC-5 fibroblasts
DFO22>25
3110.3>25
311m0.3*
ST>12.5*
NT0.5>25
N2mT>12.5>25
N4mT0.5>25
N44mT1.5>25
N4eT1.6*
N4aT2.3*
N4pT3.3*
N44pH5.2*
NoctH10.5*
FeNT>12.5*
* not determined
logPtable
3112.17
311m2.17
ST0.64
NT1.76
N2mT2.72
N4mT2.19
N44mT2.3
N4eT2.4
N4aT2.61
N4pT3.75
N44pH*
NoctH5.19
* Unable to be calculated by Broto's method
Thytable
DFO23
3110.6
311m0.6
ST>25
NT9.0
N2mT15.5
N4mT6.3
N44mT1.5
N4eT5.7
N4aT7.9
N4pT3.8
N44pH4.3
NoctH17.4
FeNT>25
mttforsem
101.9404639287110.8621252644104.914193926898.370699989292.25754223394.317077289494.8043811677100.697396073197.690160858297.2354058355128.9206537078112.8960450374101.74330310970.1
93.693476925772.002881808589.486296821382.651088282997.63388544990.439568596990.97111.306708766109.072633638690.138965084996.745117676594.386910045598.76955750860.2
40.434940399633.705492360355.891796777655.5859507208102.855019961384.295716274584.113738050798.4125710762103.767383197692.5697.327809896781.5102.62351841920.39
22.4613.773181107431.963311159926.499995097871.358426093773.682039329984.780730623990.2253281547112.957045914678.2686.812036236293.793.5352921497102.0386052855
13.13688471857.928909812716.613.409640079948.363349937651.508125656967.946427408193.256533121687.832807529962.579814247274.976100514496.188644201996.766584406892.2261964354
10.63993131436.18.47.696607573318.404687351328.33085988739.044121359250.921654156881.696716567161.681.130787089796.684.642359475896.4
9.96.678.17100853679.620264427222.838642790626.887150449211.458283012129.643362877453.678089263986.165612054475.316085860566.931285689388.6
7.84115572925.30896542935.47577989778.02972009288.68409473517.40481483217.891502760210.15569906769.958504008951.551527822264.032412254740.449390645961.896189521756.6
311
311m
NT
N4mT
N44mT
N4eT
N4aT
N4pT
N44pH
ST
N2mT
NOctH
FeNT
DFO
Chelator Concentration (mM)
Cellular Proliferation (% control)
mtttabsem
SK-N-MC neuroepithelioma
3110.3
311m0.3
NT0.5
N4mT0.5
N44mT1.5
N4eT1.6
N4aT2.3
N4pT3.3
N44pH5.2
NoctH10.5
ST>12.5
N2mT>12.5
FeNT>12.5
DFO22
mtttablesem
SK-N-MC neuroepitheliomaMRC-5 fibroblasts
3110.3>25
311m0.3>25
NT0.5>25
N4mT0.5>25
N44mT1.5>25
N2mT>12.5>25
DFO22>25
FeNT>12.5*
* not determined
NTpapMRC5sem
101.9404639287110.8621252644104.914193926898.370699989292.257542233128.92065370780.192.617449664493.048745313100.058308952294.403198175896.7415732506107.12524357060.1
93.693476925772.002881808589.486296821382.651088282997.63388544996.74511767650.289.115844762286.126334006397.1966868704103.283268587994.0952103656108.84985641520.2
40.434940399633.705492360355.891796777655.5859507208102.855019961397.32780989670.3995.885614239993.5102394001101.553934862196.251645117898.3389849717107.1040349910.39
22.4613.773181107431.963311159926.499995097871.358426093786.8120362362102.038605285583.746717245488.2030573983104.9878563539101.6428110784106.2921032183103.766943019390.2357141481
13.13688471857.928909812716.613.409640079948.363349937674.976100514492.226196435490.283046396381.7421401788107.6426712987100.0240211224103.0557669922110.325998866191.1980547746
10.63993131436.18.47.696607573318.404687351381.130787089796.466.238692734270.6662820883106.4242803477100.763901805389.3963823523111.875213361686.2899569347
9.96.678.17100853679.620264427286.165612054488.660.169244236969.743293914101.8314546437105.932619046185.6532706677105.602191372283.0391645846
7.84115572925.308965429312.58.02972009288.68409473564.032412254756.675.576305806873.665993654598.823807862397.695573033371.0267890763111.649294796480.6792129097
75.576305806865.8785.060231706492.081543233569.8859743024111.306727347672.3639224146
MRC-5Fibroblasts
SK-N-MCNeuroepithelioma
311 (S)
311m (S)
NT (S)
N4mT (S)
N44mT (S)
N2mT (S)
DFO (S)
311 (F)
311m (F)
NT (F)
N4mT (F)
N44mT (F)
N2mT (F)
DFO (F)
Chelator Concentration (mM)
Cellular Proliferation (% control)
-
EPR of ribonucleotide reductase activityControl cells311 (25mM)Triapine (25mM)
-
Mechanisms of activity - effect on cell cycle control molecules
-
Fe chelation decreases nuclear p21 proteinCONDFO 311 Cis-platinAct DMCF-7CONDFO 311 Cis-platinAct DMRC-5p53p21-actinp53p21-actinLe and Richardson (2003) Carcinogenesis 24:1045-1058NUCLEAR LYSATES
-
Decrease in nuclear p21 expression not due to cytoplasmic localizationp21CONDFO311Act Dp53MCF-7Le and Richardson (2003) Carcinogenesis 24:1045-1058
-
Gene Array Analysis
-
The increase in NDRG1 mRNA is Fe dependentNDRG1-actinCONCON + FACDFODFO + MEMDFO + FAC311 311 + MEM311 + FACMCF-7 Breast Cancer CellsTfR1
-
In Vivo Experimental DesignM109 lung carcinoma animal modelTumor weightbody weightBlood cell count, RBC, WBC, Platelet, HbOutcome assessmentTumor H&E staining and TUNEL assayTumor implantation 4Drugs injection9Stop injectionHarvesting11Day 0
-
The Role of Membrane-Bound Melanotransferrin (MTf) in Iron Uptake by Cells
-
IntercellularAdhesionScavenging Extracellular FeMetalloproteaseActivityZnMTfTfR???POSSIBLE FUNCTIONS OF MELANOTRANSFERRIN
-
Iron uptake as a function of time at 37oCand 4C for CHO cells transfected with or without MTf.
-
Effect of PI-PLC on Metal Ion Uptake by Melanoma Cells
-
E PE PMTf 2MTf 1 1 2 3 4 5 6 1 2 3 4 5 6 6b 7 8 9 10 11 12 13 14 15 16 16b MTf2 transcript ?Short MTf1 transcript (1.6 kb), exons 1 to 6 + exon 6bLong MTf1 transcript (2.3 kb) exons 1 to 16Long MTf1 transcript (2.6 kb) exons 1 to 16 + exon 16bThe relative positions of the originally identified melanotransferrin 1 (MTf1) gene compared to the newly identified MTf2 gene.
-
Total MTf Knockout Vector 5 arm3 armPGKneoTargeting VectorPGKneo5 Probe3 ProbeHomologous RecombinationWTTargetedDeletion Arm= Nhe I Restriction Enzyme site= Eco47II Restriction Site= Hind III Restriction SiteExpected Restriction Enzyme Fragments From WT and Targeted Gene5 ProbeEco47 II /Hind III double digestWT = 15 kbTargeted = 11.43 ProbeNhe I digest WT = 12.6 kbTargeted = 8.3
-
Examination of MTf Knockout MouseHistopathology/Blood Counts/Clinical Chemistry/X-ray
Morphology/Morphometry
Behaviour
Conditional Knockout