Out of more than 1100 patients treated in the p Dept of Neurosurgery, Lund University

(1)

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 för

vad som nu är Institutionen för vad som nu är Institutionen för vad som nu är Institutionen för Kliniska Vetenskaper, Lund

Kliniska Vetenskaper, Lund presenterade av hans vän

presenterade av hans vän presenterade av hans vän presenterade av hans vän och forskarkollega

och forskarkollega Leif G. Salford

Leif G. Salford

(2)

Forskningens Dag 1983 Bertil R.R. till höger i bild med hyperthermiposter ^(skymd)

(3)

GLIOMA THERAPY

THE HERBERT OLIVECRONA LECTURES

LECTURES

KAROLINSKA HOSPITAL DECEMBER 6-7 2002

(4)

(5)

Grows like an octopus.

Sends migrating

“ ill ll ”

“guerilla-cells”

into the surroun-

di l

ding normal brain

Salford

(6)

Less than 2 per thousand Less than 2 per thousand are cured from a GBM!

are cured from a GBM!

Out of more than 1100 patients treated in the p Dept of Neurosurgery, Lund University

Hospital during a 18-year period only 3 adults Hospital during a 18 year period, only 3 adults survived >10 years. They were 22, 32 and 38 years old at operation One had a recurrence years old at operation. One had a recurrence after 12 years and the other two live happily

i h d l

without tumour 42 and 38 years later.

Due to a strong immune system?

Salford 02

Due to a strong immune system?

(7)

Examples of new therapies against glioblastomas Examples of new therapies against glioblastomas

tested in the Laboratory for Experimental Neuro-Oncology:

* Magnetically responsive microspheres for drug targeting in brain tumours by the use of external magnetic gradients

* Modification of tumour cell metabolism and interference with DNA repair 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) Photodynamic therapy (PDT) with laser and hematoporphyrin derivativeswith 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)

(8)

(9)

(10)

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(12)

I t till ä kli t Inte tillräckligt bra för tumör- behandling men behandling men direkt efter den konungsliga träffen rekom- menderade prof.

P t t

Persson patents tagande för meto- dens utnyttjande dens utnyttjande som kontrastmede för MRI.

Forskningssamar- bete rivstartade

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* Whole body hyperthermia 42^oC for 30 min x 3/week for 2 weeks

* B N t C t Th (BNCT)

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Ethyl-NitrosoUrea to pregnant rat Ethyl NitrosoUrea to pregnant rat

Offspring develop transplantable CNS tumours (RG2 N32 N29)

(RG2, N32, N29)

ENU

Stereotactic implantationp

In vitro culture ^Salford^Salford

(15)

(16)

The rat glioma models RG2, N32, N29

Stereotactic implantation in

in the brain (caudate n.) in the brain (caudate n.)

In vitro culture of rat glioma cells

3 weeks later, a tumour (diam.

g

4-6 mm) giving symptoms, has developed

Salford 99 Salford 99

developed

(17)

N t Not

efficient

(18)

(19)

Cytotoxin (eg bleomycin) Cytotoxin (eg bleomycin)

not penetrating cell wall

Cell

Salford Salford

(20)

ELECTROPORATION in vivo

800 V

(21)

POST ELECTROPORATION POST ELECTROPORATION

after minutes the cell wall closes and the cytotoxin is trapped in the cell

Salford 00

(22)

Adjuvans in immunotherapy Adjuvans in immunotherapy

Opens the tumour cell wall to allow p for leakage of tumour antigens to b id tifi d b th i t

be identified by the immune system

Engström et al. Lund, Sweden

(23)

ELECTROPORATION

for immunotherapy in vivo

Immunogenic material enclosed in the tumour cell behind intact cell membrane

cell behind intact cell membrane

Salford 00

(24)

ELECTROPORATION in vivo

800 V

(25)

Spread of tumour cell content, Spread of tumour cell content,

stimulating the immune defence system

Tumour cell

Salford Salford

(26)

Experimental set up Experimental set up p p p p

Catrin Bauréus Koch Catrin Bauréus Koch

Intraperitoneal injection of

Pulse Generator for PEF treatement Intraperitoneal injection of

Radiation sterilized IFN

secreting N29 tumour cells

Contra lateral reference f

N29 tumour Impedance spectrometry

ZOK Aditus Lund Sweden

Treated Tumour^Treated

N29 tumour

(27)

(28)

Neutron capture in

¹⁰

B Neutron capture in B

4HeHe PSlh

10 m

105B n

Slh

3800 barn

(10000 ggr > än H)

Prompt gamma

7Li 5 mB

n

Thermal neutrons

(10000 ggr > än H)

Energy (2 3 MeV) Thermal neutrons

Captured by ¹⁰B Energy

Energy (2.3 MeV)

very restricted spread

(29)

N l i i ll Nuclear reaction in tumour cell

4He

10B

7Li

(30)

Bimodal radiotherapy Bimodal radiotherapy

10B ti t d

Thermal

2.

10B activated In the tumour neutrons

Inactive ¹⁰B

1.

administered intravenously

(31)

B k hi l

Boron uptake vs. histology

Histology

and boron uptake overlay

(32)

Strike in the guerilla nest Crister Ceberg et. al.

Rat glioma with a contralateral metastasis (diameter 0 2 mm) metastasis (diameter 0.2 mm) 6 hours after boron infusion.

(33)

Forskningsreaktorn i Studsvik

(34)

B t ål i iti

Bestrålningspositionen

(35)

The Rausing Laboratory for Experimental Neurosurgery and Radiation Physics, Lund University Hospital

Leif G. Salford

BRain ^Glioma I

Electro- Boron Effects

Ne tron

Chemical

Ne ro ^Stem Spinal

Bertil Persson

Immuno- Gene Tumour

Immuno Therapy.

Visse

poration and -fusion

for gene transduction

of EMF on the Brain,

BBB d

Neutron Capture Therapy

Neuro-

Anatomy ^cells

Epilepsy

Cord Injury Group

Sayer

Therapy

Translational research in

Visse Darabi Janelidze Hegardt Esbjörnsson

transduction treatment of tumours, neurological

BBB and tumour developm.

Munck af Rydelius

Al-Khaledi

collab.with dept of Tumor Immunol, 35 researchers and

Esbjörnsson Enell

g diseases

Engström Bauréus Koch

Malmgren Brun Rosensköld

Brun

y

Bexell

PI: O Nil researchers and

staff involved 5 dissert.

PI: LG Salford

2 dissert

PI:

Bauréus Koch Grafström

2 dissert

PI:

1 dissert

PI: J

2 dissert

PI: PI: G

k b

PI: J

B Nilsson

B Widegren P Siesjö B Persson C Ceberg Skagerberg Eberhardt Bengzon

(36)

1980 1980

Human IFN-

Daily localy Injections for 3 months

4 ti t 4 patients Salford Salford

Sjögren et. al.

(37)

R ti “Guerilla” ill cell

nests Brain

Resection cavity

nests

The T lymphocytes

Activation when T-lympho c tes o tside the brain are T

lympho The T-lymphocytes

must be activated in order to pass the

cytes outside the brain are presented to tumour cells which produce immune lympho

- cytes p

Blood-Brain Barrier

which produce immune stimulating products such as Interferon-

(38)

Immunisation against inoculated experimental brain tumours

N32 brain tumour cells N32 brain tumour cells

transfected with IFN-gamma genes

Day 0

Day 1 8 15

Stereotaxic

inoculation of 5000 cells

VACCINATION

Day 1,8,15

or 3,10,17

I

Day 21

DEAD CURED!

Immuneresp

(39)

BRIGTT

The 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 S

Rausing 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 G

Larsson, Karin Enell, Anita Larsson, Gunnar Gunnarsson

Tumour immunology and Bengt Widegren (Lab Chief), Hans Olov Sjögren, Genetics Xiaolong Fan Johan Rebetz Anna Edqvist

Genetics Xiaolong Fan, Johan Rebetz, Anna Edqvist

Medical Radiation Phys. Bertil Persson, Catrin Bauréus Koch, Gustaf Grafström Clinical Genetics Nils Mandahl

Neuropathology Elisabet Englund Annette Persson Arne Brun Neuropathology Elisabet Englund, Annette Persson, Arne Brun

Diagnostic Neuroradiology Elna-Marie Larsson, Cecilia Petersen, Lars Stenberg Neurology Anna Rydelius, Eva Ask

Neuropsychology Åsa Lilja, Christina Elfgren Neuropsychology Åsa Lilja, Christina Elfgren Oncology Per Malmström, Sara Kinhult

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Normal cells have stopped dividing

BRIGTT

IFN- gamma

Surgery

>80%

Resected cells 95% glioma

5% normal Normal cells divide 1/1 d

stopped dividing Glioma cells

continue and dominate the

>80%

resect.

divide 1/1 d.

Glioma cells 1/4

dominate the culture (50%)

2-3 months

Karyotyping l l

IMMUNISATION

Irradiation

regularly Lymph nodes

Activated T-cells

IMMUNISATION

3-4 months postop.

(repeated every 3 weeks,

Salford

(repeated every 3 weeks, 4 - 10 times)

(41)

Intradermal

immunisation

(42)

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)

Brain

T T

Lymph T M

M

T

nodes. T T T

T Capillary T

Blood Brain Barrier _M _T ^T

T

(43)

Brain tumour

i d i

specimen, during

immunisation period

T l h t b

T lymphocytes brown (CD3 antibody)

(44)

BRIGTT 2000-2005

Goals

:

Results:

Goals

:

Results:

to study whether vaccination with

l ll i

autologous 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)

(45)

BRIGTT Status Jan 2005 BRIGTT Status Jan 2005

# pat # immunis. age ^(op) survival after diagn

4 10 52 21 months DZ

4 10 52 y 21 months DZ

67 y 12 GF

58 y 19 5 EK

58 y 19,5 EK

64 y 24,5 IC

1 10 + 4 52 y 26 5 GA

1 10 + 4 52 y 26,5 GA 3 8 62 y 10 AMN

67 y 13 BP

67 y 13 BP

66 y 12 AO

1 6 65 y 10 KN

1 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)

(46)

BRIGTT Non-imm. pats Jan 2005 BRIGTT Non imm. pats Jan 2005

# pat # immunis. age ^(op) survival after diagn

11 0 62 8

69 6

59 5

56 6

57 17

51 17

66 6

age non-imm imm.

>60y 6,8m 13,6

66 6

59 16

66 5

<60y 12,5m 22,3 surv >18m

66 5

62 9

56 14

surv >18m

0/11 4/9

56 14

Mean age 60,3y. 9,7 months

(47)

100

Patient Survival

65 t l

80

90 65

55 58

65 62

controls patients

60 70

(%)

65 69 55

65 66

40 50

Survival (

62 61

57 67

20 30

S

50 55

63 52

57

0 10

57 59

52 63

0

0 100 200 300 400 500 600 700 800

Days after operation Age in graph

mean age controls = 59.6 age range controls = 50-69

mean age patients = 61 age range patients = 52-67

(48)

BRIGTT Future

Continue BRIGTT, add 11 patients Include patients younger than 50 y.

Utilize h-TERT cell selection? Less delay y

Add other genes? TGF-2 antisense, IL-4 etc Allogeneic cells instead of autologous? No delay Allogeneic cells instead of autologous? No delay Tumour cells direct from resected material?

C bi ith l ti i

Combine with oncolytic viruses

Combine with electropermeabilization Combine with BNCT

Combine with Stem Cell therapy py

Collaboration with other centres

(49)

MD h.c. Bertil R.R. Persson, Vår tids Carl von Linné,

(50)

Midnattsexpedition för infångande av Asplenium ruta muraria å enda dittills ruta muraria å enda dittills kända lokala lokalen.

Vem stod bakom denna Vem stod bakom denna bragd?

(51)

Bertil R R Persson Bertil R.R. Persson PhD

(52)

Bilden visar det historiska ögon historiska ögon- blicket då Bertil R.R. Persson är på väg att emot på väg att emot- taga insignierna på sin upphöjelse till MD h c

till MD h.c.

(se pilen)

(53)

Prof, general Merritt from USAF, Brooks AFBase, San Antonio

(54)

EMF – BBB studier startade 1988 vid Lab. för Exp. Neuroonkologi, USiL

(55)

Världens största biologiska experiment Världens största biologiska experiment 25% av jordens befolkning frivilliga

25% av jordens befolkning frivilliga försöks

försöks kaniner kaniner

försöks

försöks -- kaniner kaniner

(56)

On-line publication 20030129

20030129

I i t J 2003

In print June 2003

(57)

10

Em d( ) 7.75

meter

SAR

3.25 5.5

Volt/m

d

0 0.5 1 1.5 2

1

SAR=1mW/kg

d

meter ^{1.85 meter}

SAR = 1mW/kg 1.85 metres away

f th bil

from the mobile phone

d

(58)

1010

Eb d( ) 7.75

meter

Eb d( ) 7.75

meter

( ) SAR

3.25 5.5

Volt/m ^{( )}

SAR

3.25 5.5

Volt/m

0 50 100 150 200 250

1 SR=1mW/kg

0 50 100 150 200 250

1 SAR=1mW/kg

d meter

190 meter d

meter

190 meter

(59)



?  NMT

NMT GSM GSM 3G 3G

?  CW

CW 217 Hz 217 Hz ^Different ^Different

microwaves!

The Lund group has shown in a large material that The 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 sends

in the RAT brain. The 3G technique sends

microwaves of a different character, and it is quite possible that the biological effect of 3G differs from possible that the biological effect of 3G differs from that of NMT och GSM.

We want to test this hypothesis in our animal We want to test this hypothesis in our animal model.

(60)

TACK BERTIL !

Polfarare också i de Polfarare också i de magnetiska fälten

Vännen Leif

(61)

O k å IKVL ill t k i hö t

Också 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.)

Institutionens för Kliniska Vetenskapers samtliga 40 avdelningar tackar

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