några glimtar ur professor bertil r.r. perssons ... · några glimtar ur professor bertil r.r....
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Några glimtar ur professor Bertil R.R. Perssons Några glimtar ur professor Bertil R.R. Perssons translationella gärning under 4 decennier vid translationella gärning under 4 decennier vid vad som nu är Institutionen förvad som nu är Institutionen förvad som nu är Institutionen förvad som nu är Institutionen förKliniska Vetenskaper, LundKliniska Vetenskaper, Lund
presenterade av hans vänpresenterade av hans vänpresenterade av hans vänpresenterade av hans vänoch forskarkollegaoch forskarkollegaLeif G. SalfordLeif G. Salford
Forskningens Dag 1983 Bertil R.R. till höger i bild med hyperthermiposter (skymd)
GLIOMA THERAPY
THE HERBERT OLIVECRONA LECTURESLECTURESKAROLINSKA HOSPITALDECEMBER 6-7 2002
Grows like anoctopus.Sends migrating“ ill ll ”“guerilla-cells”into the surroun-di lding normal brain
Salford
Less than 2 per thousandLess than 2 per thousandare cured from a GBM!are cured from a GBM!
Out of more than 1100 patients treated in the pDept of Neurosurgery, Lund University Hospital during a 18-year period only 3 adultsHospital during a 18 year period, only 3 adultssurvived >10 years. They were 22, 32 and 38years old at operation One had a recurrenceyears old at operation. One had a recurrence after 12 years and the other two live happily
i h d lwithout tumour 42 and 38 years later.Due to a strong immune system?
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Due to a strong immune system?
Examples of new therapies against glioblastomasExamples of new therapies against glioblastomastested in the Laboratory for Experimental Neuro-Oncology:
* Magnetically responsive microspheres for drug targeting in braintumours by the use of external magnetic gradients
* Modification of tumour cell metabolism and interference with DNArepair by Chlorpromazine or Methoclopramide and BCNU
* M difi ti f th li htl lk li t li H f t ll b* Modification of the slightly alkaline cytosolic pH of tumour cells by the use of salicylate
* Photodynamic therapy (PDT) with laser and hematoporphyrin derivatives Photodynamic therapy (PDT) with laser and hematoporphyrin derivatives* Radioactive Thallium (Tl 201) which accumulates in the tumour cells* Electroporation in vivo in the brain parenchyma with or without
concomitant hydrophilic cytotoxin treatment (e.g. Bleomycin)* Boron Neutron Capture Therapy (BNCT)
I t till ä kli tInte tillräckligtbra för tumör-behandling menbehandling mendirekt efter denkonungsliga träffen rekom-menderade prof.P t tPersson patentstagande för meto-dens utnyttjandedens utnyttjandesom kontrastmedeför MRI.Forskningssamar-bete rivstartade
Examples of new therapies against glioblastomasExamples of new therapies against glioblastomastested in the Laboratory for Experimental Neuro-Oncology:
* Magnetically responsive microspheres for drug targeting in braintumours by the use of external magnetic gradients
* Modification of tumour cell metabolism and interference with DNArepair by Chlorpromazine or Methoclopramide and BCNU
* M difi ti f th li htl lk li t li H f t ll b* Modification of the slightly alkaline cytosolic pH of tumour cells by the use of salicylate
* Photodynamic therapy (PDT) with laser and hematoporphyrin derivatives Photodynamic therapy (PDT) with laser and hematoporphyrin derivatives* Radioactive Thallium (Tl 201) which accumulates in the tumour cells* Whole body hyperthermia 42oC for 30 min x 3/week for 2 weeks* Electroporation in vivo in the brain parenchyma with or without
concomitant hydrophilic cytotoxin treatment (e.g. Bleomycin)* B N t C t Th (BNCT)* Boron Neutron Capture Therapy (BNCT)
Ethyl-NitrosoUrea to pregnant ratEthyl NitrosoUrea to pregnant ratOffspring develop transplantable CNS tumours
(RG2 N32 N29)(RG2, N32, N29)ENU
Stereotacticimplantationp
In vitro culture SalfordSalford
The rat glioma models RG2, N32, N29Stereotacticimplantation in in the brain (caudate n.)in the brain (caudate n.)
In vitro cultureof rat glioma cells
3 weeks later,a tumour (diam. g4-6 mm) giving symptoms, has developed
Salford 99Salford 99
developed
N tNot efficient
Examples of new therapies against glioblastomasExamples of new therapies against glioblastomastested in the Laboratory for Experimental Neuro-Oncology:
* Magnetically responsive microspheres for drug targeting in braintumours by the use of external magnetic gradients
* Modification of tumour cell metabolism and interference with DNArepair by Chlorpromazine or Methoclopramide and BCNU
* M difi ti f th li htl lk li t li H f t ll b* Modification of the slightly alkaline cytosolic pH of tumour cells by the use of salicylate
* Photodynamic therapy (PDT) with laser and hematoporphyrin derivatives Photodynamic therapy (PDT) with laser and hematoporphyrin derivatives* Radioactive Thallium (Tl 201) which accumulates in the tumour cells* Electroporation in vivo in the brain parenchyma with or without
concomitant hydrophilic cytotoxin treatment (e.g. Bleomycin)* Boron Neutron Capture Therapy (BNCT)
Cytotoxin (eg bleomycin)Cytotoxin (eg bleomycin)not penetrating cell wall
Cell
SalfordSalford
ELECTROPORATION in vivo800 V
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POST ELECTROPORATIONPOST ELECTROPORATIONafter minutes the cell wall closes and the cytotoxin is trapped in the cell
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Adjuvans in immunotherapyAdjuvans in immunotherapyOpens the tumour cell wall to allow p
for leakage of tumour antigens to b id tifi d b th i tbe identified by the immune system
Engström et al. Lund, Sweden
ELECTROPORATION for immunotherapy in vivoImmunogenic material enclosed in the tumourcell behind intact cell membranecell behind intact cell membrane
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ELECTROPORATION in vivo800 V
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Spread of tumour cell content,Spread of tumour cell content, stimulating the immune defence system
Tumour cell
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Experimental set upExperimental set upp pp pCatrin Bauréus KochCatrin Bauréus Koch
Intraperitoneal injection of Pulse Generator forPEF treatementIntraperitoneal injection of
Radiation sterilized IFNsecreting N29 tumour cells
Contra lateral reference fN29 tumour Impedance spectrometry
ZOK Aditus Lund SwedenTreated TumourTreated N29 tumour
Examples of new therapies against glioblastomasExamples of new therapies against glioblastomastested in the Laboratory for Experimental Neuro-Oncology:
* Magnetically responsive microspheres for drug targeting in braintumours by the use of external magnetic gradients
* Modification of tumour cell metabolism and interference with DNArepair by Chlorpromazine or Methoclopramide and BCNU
* M difi ti f th li htl lk li t li H f t ll b* Modification of the slightly alkaline cytosolic pH of tumour cells by the use of salicylate
* Photodynamic therapy (PDT) with laser and hematoporphyrin derivatives Photodynamic therapy (PDT) with laser and hematoporphyrin derivatives* Radioactive Thallium (Tl 201) which accumulates in the tumour cells* Electroporation in vivo in the brain parenchyma with or without
concomitant hydrophilic cytotoxin treatment (e.g. Bleomycin)* Boron Neutron Capture Therapy (BNCT)
Neutron capture in 10BNeutron capture in B
4He SlhPHe10 m
510Bn
Slh3800 barn
(10000 ggr > än H)
Prompt gamma
7Li
5 mBn
Thermal neutrons
(10000 ggr > än H)
Energy (2 3 MeV)
Thermal neutronsCaptured by 10B Energy
Energy (2.3 MeV)very restricted spread
N l i i llNuclear reaction in tumour cell
4He
10B7Li7Li
Bimodal radiotherapyBimodal radiotherapy
10B ti t dThermal 2.10B activatedIn the tumourneutrons
Inactive 10B 1.
administeredintravenously
B k hi lBoron uptake vs. histology
Histology
and boron uptake
overlay
Strike in the guerilla nestCrister Ceberg et. al.
Rat glioma with a contralateralmetastasis (diameter 0 2 mm)metastasis (diameter 0.2 mm)6 hours after boron infusion.
Forskningsreaktorn i Studsvik
B t ål i itiBestrålningspositionen
The Rausing Laboratory for Experimental Neurosurgeryand Radiation Physics, Lund University Hospital
Leif G. Salford
BRain GliomaI
Electro- EffectsBoronNe tron
ChemicalNe ro Stem Spinal
Bertil Persson
Immuno-GeneTumour
ImmunoTherapy.
Visse
porationand -fusionfor genetransduction
of EMFon theBrain, BBB d
NeutronCaptureTherapy
Neuro-Anatomy
cells
Epilepsy
CordInjuryGroup
Sayer
TherapyTranslationalresearch in
VisseDarabiJanelidzeHegardtEsbjörnsson
transductiontreatment oftumours,neurological
BBB andtumourdevelopm.
Munck af RydeliusAl-Khaledicollab.with
dept of Tumor Immunol, 35 researchers and
EsbjörnssonEnell
gdiseases
EngströmBauréus Koch
MalmgrenBrun
RosensköldBrun
y
Bexell
PI: ONil
researchers and staff involved5 dissert.PI: LG Salford
2 dissertPI:
Bauréus KochGrafström2 dissertPI:
1 dissert
PI: J
2 dissert
PI: PI: Gk b
PI: JB NilssonB Widegren P Siesjö B Persson EberhardtC Ceberg Skagerberg Bengzon
19801980Human IFN-Daily localyInjections for3 months4 ti t4 patients
SalfordSalfordSjögren et. al.
illR ti “Guerilla” cellnests
BrainResectioncavity
nests
The T lymphocytesActivation when T-lymphoc tes o tside the brain are
T lympho
The T-lymphocytesmust be activated in order to pass the
cytes outside the brain are presented to tumour cellswhich produce immunelympho
- cytesp
Blood-Brain Barrierwhich produce immune stimulating products such as Interferon-
Immunisation against inoculated experimental brain tumours
N32 brain tumour cells N32 brain tumour cellstransfected with IFN-gamma genes
Day 0Day 1 8 15
Stereotaxicinoculation of 5000 cells
VACCINATION
Day 1,8,15or 3,10,17
I
Day 21 DEAD CURED!Immuneresp
BRIGTTThe Brain Immuno Gene Tumour Therapy project The Rausing Laboratory, Lund University and Lund University Hospital,
Lund, Sweden
Neurosurgery and the Leif G. Salford (PI), Peter Siesjö, Gunnar Skagerberg, R i l b C li L Ch l O SRausing laboratory Carolina Larsson, Charlotte Orre, Susanne
Strömblad, Catarina Blennow, Lena-Liebera-Jannert, Edward Visse, Shorena Janelidze, Anna Darabi, Maria L K i E ll A it L GLarsson, Karin Enell, Anita Larsson, Gunnar Gunnarsson
Tumour immunology and Bengt Widegren (Lab Chief), Hans Olov Sjögren, Genetics Xiaolong Fan Johan Rebetz Anna EdqvistGenetics Xiaolong Fan, Johan Rebetz, Anna EdqvistMedical Radiation Phys. Bertil Persson, Catrin Bauréus Koch, Gustaf GrafströmClinical Genetics Nils Mandahl Neuropathology Elisabet Englund Annette Persson Arne BrunNeuropathology Elisabet Englund, Annette Persson, Arne BrunDiagnostic Neuroradiology Elna-Marie Larsson, Cecilia Petersen, Lars StenbergNeurology Anna Rydelius, Eva AskNeuropsychology Åsa Lilja, Christina ElfgrenNeuropsychology Åsa Lilja, Christina ElfgrenOncology Per Malmström, Sara Kinhult
Normal cells havestopped dividing
BRIGTTIFN-gamma
Surgery>80%
Resected cells95% glioma 5% normal Normal cells
divide 1/1 d
stopped dividingGlioma cellscontinue anddominate the>80%
resect.divide 1/1 d. Glioma cells 1/4
dominate theculture (50%)
2-3 monthsKaryotyping
l l
IMMUNISATIONIrradiation
regularly
Lymph nodesActivated T-cells
IMMUNISATION3-4 months postop.
(repeated every 3 weeks,
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(repeated every 3 weeks,4 - 10 times)
IntradermalIntradermal immunisation
Gene-modified brain tumour cells for immunisation against malignant brain tumours (the BRIGTT study)
Antigen-specific T-cells (T) pass actively through the BBB and localize the migrated “guerilla” cells. (M: monocytes)
BrainT T
TT TT
TTT M
LymphM
T
T
T Tnodes. T
T
CapillaryTBlood Brain Barrier T
M TT
T
Brain tumouri d ispecimen, during
immunisation periodT l h t bT lymphocytes brown (CD3 antibody)
BRIGTT 2000-2005 Goals: Results:Goals: Results:to study whether vaccination with
l ll iautologous tumour cells expressing gene-sequences for human interferon-gamma 1 is safe for the patients YES !1. is safe for the patients YES !2. gives rise to an immunological
YES !response YES !3. adds any beneficial effect to
conventional therapy (tumor growth, ?l d i l)prolonged survival)
BRIGTT Status Jan 2005BRIGTT Status Jan 2005# pat # immunis. age (op) survival after diagn
4 10 52 21 months DZ4 10 52 y 21 months DZ67 y 12 GF58 y 19 5 EK58 y 19,5 EK64 y 24,5 IC
1 10 + 4 52 y 26 5 GA1 10 + 4 52 y 26,5 GA3 8 62 y 10 AMN
67 y 13 BP67 y 13 BP66 y 12 AO
1 6 65 y 10 KN1 6 65 y 10 KN
Mean age 61 4y Mean survival time 16 4 m (169%)Mean age 61,4y Mean survival time 16,4 m (169%)(c.f. not imm. pats 60,3y 9,7 m)
BRIGTT Non-imm. pats Jan 2005BRIGTT Non imm. pats Jan 2005# pat # immunis. age (op) survival after diagn
11 0 62 811 0 62 869 659 559 556 657 1757 1751 1766 6
age non-imm imm.>60y 6,8m 13,6
66 659 1666 5
<60y 12,5m 22,3
surv >18m66 562 956 14
surv >18m0/11 4/9
56 14Mean age 60,3y. 9,7 months
100Patient Survival
65 t l
80
9065
55
5865
62
controlspatients
60
70
(%) 65
69
55
65
66
40
50
Sur
viva
l (
62
61
57
67
20
30
S
50
55
63
52
57
0
1057
5952
63
00 100 200 300 400 500 600 700 800
Days after operationAge in graphmean age controls = 59.6age range controls = 50-69
mean age patients = 61age range patients = 52-67
BRIGTT FutureBRIGTT Future Continue BRIGTT, add 11 patientsInclude patients younger than 50 y.Utilize h-TERT cell selection? Less delayyAdd other genes? TGF-2 antisense, IL-4 etcAllogeneic cells instead of autologous? No delayAllogeneic cells instead of autologous? No delay Tumour cells direct from resected material?C bi ith l ti iCombine with oncolytic virusesCombine with electropermeabilizationCombine with BNCTCombine with Stem Cell therapypyCollaboration with other centres
MD h.c. Bertil R.R. Persson, Vår tids Carl von Linné,
Midnattsexpedition för infångande av Aspleniumruta muraria å enda dittillsruta muraria å enda dittillskända lokala lokalen.
Vem stod bakom dennaVem stod bakom dennabragd?
Bertil R R PerssonBertil R.R. PerssonPhD
Bilden visar det historiska ögonhistoriska ögon-blicket då BertilR.R. Persson är på väg att emotpå väg att emot-taga insigniernapå sin upphöjelsetill MD h ctill MD h.c.(se pilen)
Prof, general Merritt from USAF, Brooks AFBase, San AntonioProf, general Merritt from USAF, Brooks AFBase, San Antonio
EMF – BBB studier startade 1988 vid Lab. för Exp. Neuroonkologi, USiL
© Salford et al
Världens största biologiska experimentVärldens största biologiska experiment25% av jordens befolkning frivilliga25% av jordens befolkning frivilligaförsöksförsöks kaninerkaninerförsöks försöks -- kaniner kaniner
On-line publication 2003012920030129
I i t J 2003In print June 2003
10
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d0 0.5 1 1.5 2
1SAR=1mW/kg
dmeter 1.85 meter
SAR = 1mW/kg1.85 metres awayf th bilfrom the mobilephone
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0 50 100 150 200 2501
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SAR=1mW/kg
dmeter
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d © Salford et al
?NMTNMT GSMGSM 3G3G
?
CWCW 217 Hz217 Hz DifferentDifferentmicrowaves!microwaves!microwaves!microwaves!
The Lund group has shown in a large material thatThe Lund group has shown in a large material that both NMT and GSM open the Blood-Brain Barrier in the RAT brain The 3G technique sendsin the RAT brain. The 3G technique sends microwaves of a different character, and it is quite possible that the biological effect of 3G differs frompossible that the biological effect of 3G differs from that of NMT och GSM. We want to test this hypothesis in our animalWe want to test this hypothesis in our animal model.
© Salford et al
TACK BERTIL !Polfarare också i dePolfarare också i demagnetiska fälten
Vännen Leif
O k å IKVL ill t k i hö tOckså IKVL vill tacka sin oerhört upp-skattade professor sen många decen-
nier Bertil R.R. Persson, MD h.c.(med en omöjlig figur av O R v g v )(med en omöjlig figur av O.R. v.g.v.)
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