Proteinases in Inflammation and Tumor Invasion Review Articles including those from an International Conference Bielefeld, Federal Republic of Germany March 14-16,1985

Editor Harald Tschesche

W DE G

Walter de Gruyter • Berlin • New York 1986

Editor Harald Tschesche, Dr. rer. nat. Professor für Biochemie Lehrstuhl für Biochemie Fakultät für Chemie Universität Bielefeld D-4800 Bielefeld Federal Republic of Germany

Universitäts-Bibliothek München

Library of Congress Cataloging in Publication Data

Proteinases in inflammation and tumor invasion

Includes bibliographies and index. 1. Inflammation—Congresses. 2. Proteinase—Physiological effect—Congresses. Cancer invasiveness— Congresses. 4. Collagen diseases—Congresses. I. Tschesche, Harald. [DNLM: 1. Inflammation—congresses. 2. Neoplasm Invasiveness—congresses. 3. Peptide Peptidohydrolases—congresses. QZ 202 P967 1985] RB131.P77 1986 616'.0473 86-16708 ISBN 0-89925-216-8 (U.S.)

CIP-Kurztitelaufnahme der Deutschen Bibliothek

Proteinases in inflammation and tumor invasion : review articles including those from an international conference, Bielefeld, Fed. Republic of Germany, March 14 -16,1985 / ed. Harald Tschesche. - Berlin ; New York : de Gruyter, 1986.

ISBN 3-11-010530-6

NE: Tschesche, Harald [Hrsg.]

Copyright © 1986 by Walter de Gruyter & Co., Berlin 30. All rights reserved, including those of translation into foreign languages. No part of this book may be reproduced in any form - by photoprint, microfilm or any other means nor transmitted nor translated into a machine language without written permission from the publisher. Printing: Gerike GmbH, Berlin. - Binding: D. Mikolai, Berlin. - Printed in Germany.

CONTENTS

G r a n u l o c y t e P r o t e i n a s e s as M e d i a t o r s o f U n s p e c i f i c P r o t e o l y s i s i n I n f l a m m a t i o n : A R e v i e w

H. F r i t z , M. J o c h u m , K.H. D u s w a l d , H. D i t t m e r , H. K o r t m a n n , S. Neumann, H. Lang 1

E f f e c t o f G r a n u l o c y t e E l a s t a s e on t h e M e t a b o l i s m o f P r o t e o g l y c a n s i n I n f l a m m a t o r y J o i n t D i s e a s e s

K. K l e e s i e k , R. R e i n a r d s , H. G r e i l i n g 25

P r o t e i n a s e I n h i b i t o r T h e r a p y o f S e v e r e I n f l a m m a t i o n i n P i g s : F i r s t R e s u l t s w i t h E g ! i n , a P o t e n t I n h i b i t o r o f G r a n u l o c y t e E l a s t a s e and C a t h e p s i n G

M. J o c h u m , H.F. W e l t e r , M. S i e b e c k , H. F r i t z 53

Mast C e l l s and M a t r i x D e g r a d a t i o n i n t h e R h e u m a t o i d J o i n t

D.E. W o o l l e y , J.R. Y o f f e , J.M. E v a n s o n 6 1

Human C a t h e p s i n B, H and L and t h e i r E n d o g e n o u s P r o t e i n I n h i b i t o r s : S t r u c t u r e , F u n c t i o n and B i o l o g i c a l R o l e

V. T u r k , J . B r z i n , M. K o p i t a r , M. K o t n i k , B. L e n a r c i c , T. P o p o v i c , A. R i t o n j a , M. T r s t e n j a k , B. Rozman, W. M a c h l e i d t 77

C a t h e p s i n B: A P r o t e i n a s e L i n k e d t o M e t a s t a s i s ?

B. F. S l o a n e , R.E. R y a n , J . R o z h i n , T.T. L a h , J.D. C r i s s m a n , K. V. Honn 93

The C o l l a g e n s - M o l e c u l a r and M a c r o m o l e c u l a r S t r u c t u r e s

K. Kühn 107

P r o t e o l y t i c M o d i f i c a t i o n s o f T y p e V I C o l l a g e n

J . R a u t e r b e r g , R. J a n d e r , D. T r o y e r 145

D e g r a d a t i o n o f S p e c i f i c C o l l a g e n s by M a c r o p h a g e s and N e u t r o p h i 1 s

C. L. M a i n a r d i , M.S. H i b b s , K.A. H a s t y 161

C o n n e c t i v e T i s s u e M e t a l 1 o p r o t e i n a s e s and M a t r i x D e g r a d a t i o n

G. M u r p h y , J . G a v r i l o v i c , Ch. M c A l p i n e 173

VIII

T i s s u e I n h i b i t o r o f M e t a 1 1 o p r o t e i n a s e s

T.E. C a w s t o n , E. M e r c e r , C. Mooney, J.K. W r i a h t 189

C o l l a g e n D e g r a d a t i o n i n P e r i o d o n t a l D i s e a s e s

V . - J . U i t t o 211

I n t e r s t i t i a l C o l l a g e n a s e , G e l a t i n a s e and a S p e c i f i c T y p e I V / V ( B a s e m e n t Membrane) C o l l a g e n D e g r a d i n q P r o t e i n a s e f r o m Human L e u k o c y t e s

H. T s c h e s c h e , J . F e d r o w i t z , U. K o h n e r t , H.W. M a c a r t n e y , J . M i c h a e l i s , K. K ü h n , H. Wiedemann 2 2 5

The R o l e o f C e l l M o t i l i t y i n Tumor I n v a s i o n

G. H a e m m e r l i 2 4 5

P e r i t u m o r a l E x t r a c e l l u l a r M a t r i x D e g r a d a t i o n i n t h e L o o s e C o n n e c t i v e T i s s u e o f t h e M e s e n t e r y

W. M ü l 1 e r - G l a u s e r 2 6 1

P l a s m i n o g e n A c t i v a t o r s and Tumor I n v a s i o n

L. S k r i v e r , L.R. L u n d , P.A. A n d r e a s e n , P. K r i s t e n s e n , K. Dan0 2 8 1

The R o l e o f P r o t e i n a s e I n h i b i t o r s i n C e l l u l a r I n v a s i v e n e s s : P u r i f i c a t i o n and C h a r a c t e r i z a t i o n o f a S e r i n e P r o t e i n a s e I n h i b i t o r f r o m R a t C h o n d r o s a r c o m a

H. L e v y , G. F e i n s t e i n 3 0 5

C h a r a c t e r i z a t i o n o f a C o l l a g e n a s e I n h i b i t o r f r o m Human A m n i o t i c F l u i d w h i c h I n h i b i t s Tumor C e l l I n v a s i o n

H. L e v y , P. M i g n a t t i , D.B. R i f k i n 3 2 1

D e v e l o p m e n t o f a D i r e c t M e a s u r e m e n t A s s a y f o r C o l l a g e n a s e a g a i n s t Type I and Type IV C o l l a g e n s i n T i s s u e H o m o g e n a t e and i t s A p p l i c a t i o n i n Stomach and Lung C a n c e r s

K. K u b o c h i , K. Maruyama, I . O k a z a k i 3 3 7

M e t a s t a t i c A b i l i t y o f L e w i s Lung C a r c i n o m a C e l l s and E x t r a c e l l u l a r P r o t e o l y s i s . I n t e r a c t i o n s b e t w e e n Tumour C e l l s and H o s t M a c r o p h a g e s i n t h e D e g r a d a t i o n o f Type I and T y p e IV C o l l a g e n

G. V a e s , N. H e n r y , A.L. van L a m s w e e r d e , Y. E e c k h o u t 3 5 7

IX

A T-Lymphoma D e r i v e d P r o t e i n a s e , S y n e r g i z i n g w i t h an E n d o g l y c o s i d a s e i n t h e D e g r a d a t i o n o f S u l p h a t e d P r o ­t e o g l y c a n s i n S u b e n d o t h e l i a l E x t r a c e l l u l a r M a t r i x

M.D. K r a m e r , V. S c h i r r m a c h e r , M. B a r - N e r , I . V l o d a v s k y 373

P r o t e o l y t i c A c t i v i t y i n C u l t u r e d Human Lung C a n c e r C e l l s

O.L. S c h o e n b e r q e r , H. S c h w ö b e l , W. E b e r t , P. A u l e n b a c h e r , H.O. W e r l i n g , N. P a w e l e t z 395

L i s t o f A u t h o r s 409

S u b j e c t I ndex 411

PROTEINASE INHIBITOR THERAPY OF SEVERE INFLAMMATION IN PIGS: FIRST RESULTS WITH EGLIN, A POTENT INHIBITOR OF GRANULOCYTE ELASTASE AND CATHEPSIN G

M. Jochum, H. F. We l t e r * , M. Siebeck* and H. F r i t z

Dept. o f C l i n i c a l C h e m i s t r y and C l i n i c a l B i o c h e m i s t r y i n t h e *Surgery C l i n i c C i t y , U n i v e r s i t y of Munich, Munich, FRG

I n t r o d u c t i o n

Lysosomal enzymes r e l e a s e d d u r i n g i n f l a m m a t i o n may enhance the i n f l a m m a t o r y response by s p e c i f i c a c t i v a t i o n of humoral systems ( c l o t t i n g , f i b r i n o l y s i s , complement, k a l l i k r e i n / k i n i n system) o r by u n s p e c i f i c d e g r a d a t i o n and o x i d a t i o n o f plasma f a c t o r s and t i s s u e p r o t e i n s . R e c e n t l y , the pa t h o b i o c h e m i c a l e f f e c t s of enzymes of the a z u r o p h i l i c and s p e c i f i c g r a n u l e s o f polymorpho­n u c l e a r g r a n u l o c y t e s ( e l a s t a s e , c a t h e p s i n G, collagenase, ca­t h e p s i n B, lysozyme, myeloperoxidase etc.) were i n v e s t i g a t e d i n more d e t a i l ( f o r r e v i e w o f l i t e r a t u r e see ( 1 ) ) . N o r m a l l y , t h e p r o t e o l y t i c a c t i o n o f the lysosomal p r o t e i n a s e s i s balanced by p r o t e i n a s e i n h i b i t o r s p r e s e n t i n plasma or o t h e r body f l u i d s . D uring severe i n f l a m m a t i o n , however, these p r o t e i n a s e s may be l i b e r a t e d i n amounts exceeding the n a t u r a l i n h i b i t o r l e v e l s so t h a t d e s t r u c t i v e processes can occur e i t h e r l o c a l l y or system-i c a l l y . Evidence c o u l d be presented t h a t l e u k o c y t i c e l a s t a s e i s indeed r e l e a s e d d u r i n g s e p t i c e m i a and o t h e r i n f l a m m a t o r y pro­cesses ( 1 ) . The c o r r e l a t i o n o b s e r v e d between t h e r e l e a s e o f t h i s p r o t e i n a s e and the consumption of plasma f a c t o r s i n d i c a t e s t h a t l y s o s o m a l enzymes may s i g n i f i c a n t l y c o n t r i b u t e t o t h e consumption of plasma f a c t o r s by u n s p e c i f i c p r o t e o l y s i s . Fur-t h e r o n , a d m i n i s t r a t i o n of an exogenous i n h i b i t o r of lysosomal p r o t e i n a s e s i n an e x p e r i m e n t a l a n i m a l model ( e n d o t o x e m i a ) caused a s i g n i f i c a n t r e d u c t i o n o f t h e c o n s u m p t i o n o f plasma f a c t o r s ( 2 ) .

Proteinases in Inflammation and Tumor Invasion © 1986 Walter de Gruyter & Co., Berlin • New York - Printed in Germany

54

E g l i n — a p r o t e i n a s e i n h i b i t o r which o r i g i n a l l y has been i s o ­l a t e d from t h e l e e c h Hirudo m e d i c i n a l i s — i s a p o t e n t i n h i b i t o r of the g r a n u l o c y t e p r o t e i n a s e s e l a s t a s e and c a t h e p s i n G ( 3 ) . With regard t o t h e r o l e o f these p r o t e i n a s e s i n i n f l a m m a t o r y s t a t e s l i k e s e p t i c shock and trauma e v a l u a t i o n o f a t h e r a p e u t i c approach by p r o t e i n a s e i n h i b i t i o n seemed t o be d e s i r a b l e . Since e g l i n i s now a v a i l a b l e i n s u f f i c i e n t q u a n t i t i e s (Fa. Ciba Geigy/Basel) by g e n e t i c e n g i n e e r i n g ( 4 ) e x p e r i m e n t a l animal s t u d i e s were performed i n o r d e r t o e l u c i d a t e the i n h i b i t o r y potency of t h i s i n h i b i t o r i n sepsis and s e p t i c shock.

Methods

Septicemia was induced i n p i g s by i . v . i n f u s i o n of E . c o l i f o r 2 hours ( 5 ) . Three d i f f e r e n t p r e p a r a t i o n s of e g l i n were i n ­t r a v e n o u s l y a d m i n i s t e r e d f o r 4 hours: cloned e g l i n c ( b a t c h I or batch I I ) o r e g l i n b ( i s o l a t e d from the l e e c h ) . Animals (16.5 - 23 kg) were d i v i d e d i n t o 7 groups (Tab. 1 ) .

Hematologic, hemodynamic, r e s p i r a t o r y , h i s t o l o g i c a l and bioche­m i c a l parameters were measured by r o u t i n e methods; e x t r a v a s c u -l a r lung water (EVLW) was e v a l u a t e d u s i n g the thermo-dye-technique ( 6 ) . E g l i n l e v e l s i n plasma and u r i n e were e s t i m a t e d by r a d i a l i m m u n o d i f f u s i o n a p p l y i n g s p e c i f i c a n t i b o d i e s ( 7 ) .

Results

Pharmacokinetic d a t a : E g l i n was e x c r e t e d i n u r i n e t o 75-95 % w i t h i n 12 h i n animals w i t h normal kidney f u n c t i o n . Animals w i t h r e n a l i n s u f f i c i e n c y due t o s e p t i c e m i a showed a s i g n i f i c a n t accumulation of e g l i n i n the c i r c u l a t i o n i n d i c a t i n g the p r e ­v a i l i n g r e n a l e l i m i n a t i o n of t h i s i n h i b i t o r .

55

S u r v i v a l time (Tab. 1 ) : Animals of group I I and I I I d i e d w i t h i n 7.2 _+ 3.0 h and 5.3 _+ 1.5 h, r e s p e c t i v e l y . One p i g o f each of these two groups s u r v i v e d 20 hours. Four animals o f group IV s u r v i v e d an e x p e r i m e n t a l p e r i o d o f 30 h d e s p i t e f e v e r due t o t r a c e s o f e n d o t o x i n p r e s e n t i n batch I o f t h e c l o n e d e g l i n c. The mean s u r v i v a l time i n group V was 19.1 + 6 h. I n group VI and V I I e n d o t o x i n - f r e e e g l i n was used and a remarkable p r o l o n ­g a t i o n o f s u r v i v a l time was observed i n s p i t e o f t y p i c a l hema­t o l o g i c a l , hemodynamic and r e s p i r a t o r y d i s o r d e r s due t o s e p t i ­cemia .

Group B a c t e r i a dosage ( E . c o l i c e l l s )

E g l i n dosage (mg/kg x h)

S u r v i v a l time

No n i n 12 ml phys. NaCl/h f o r 2 h

i n 50 ml phys.NaCl/h f o r 4 h

h

I 10 - - 30

I I 8 3 x 10 9 - 7.2 + 3.0

I I I 9 3 x 1 0 1 0 - 5.3 + 1 .5

IV 5 - 3.85 ( E g l i n c; charge I )

3 0 ( 4 x ) ; 6. 5(1x)

V 5 3 x 1 0 9 3.85 ( E g l i n c; charge I )

19.1 + 9.6

VI 2 3 x 10 9 4.55 ( E g l i n b)

3 0 ( 1 x ) ; 4. 5(1x)

V I I 7 3 x 1 0 1 0 3.85 ( E g l i n c; charge I I )

30(5x);15. 2(2x)

Table 1: S u r v i v a l time o f p i g s i n e x p e r i m e n t a l bacteremia as a f u n c t i o n of b a c t e r i a ( E . c o l i ) and i n h i b i t o r ( E g l i n ) dosages.

56

Lung d i s o r d e r s : A s i g n i f i c a n t i n c r e a s e i n e x t r a v a s c u l a r lung water was measured i n u n t r e a t e d s e p t i c a n i m a l s , whereas d u r i n g and a f t e r e g l i n - a d m i n i s t r a t i o n o n l y a minor r i s e i n EVLW oc-cured ( F i g , 1 ) . Moreover, i n c o n t r a s t t o u n t r e a t e d s e p t i c p i g s e g l i n - t r e a t e d animals showed o n l y modest m o r p h o l o g i c a l lung a l t e r a t i o n s ( i n t e r s t i t i a l edema).

•= i t 7 • 7 4 3 2 I I I I I » 1 * * ? ? ? 1 ? ? ? ? l ? ? ? 7 7 l

• -—I , — c • t C C t t t C t t t * • L t t t t C t t t 1 J » • - «

F i g . 1: Mean e x t r a v a s c u l a r lung water l e v e l s (EVLW) i n s e p t i c p i g s w i t h o u t (a) and w i t h i n h i b i t o r (b) t r e a t m e n t .

C l o t t i n g f a c t o r s : I n the e g l i n - t r e a t e d groups I V - V I I f a c t o r X I I I and a n t i t h r o m b i n I I I l e v e l s decreased t r a n s i e n t l y ( F i g . 2b, 3b), whereas i n the u n t r e a t e d bacteremic animals (group I I and I I I ) a s i g n i f i c a n t consumption of f a c t o r X I I I ( F i g . 2a) and a n t i t h r o m b i n I I I ( F i g . 3a) was measured u n t i l death.

57

a Bacteremia

E.coii 3 • i m

1

s

L * S 1 7 4 4 2 4 • 1 • I # » »

Bacteremia * Eglin E.cli 3 i i m - Efll. 3.95 t ^ t U

T-H, I TT"

: S 5 $ 5 S 5 5 5 • 5 • 5 • $ 4 J

* * " * *Ttat (Urn)

F i g . 2: Mean plasma l e v e l s of f a c t o r X I I I i n s e p t i c p i g s w i t h ­out (a) and w i t h i n h i b i t o r (b) t r e a t m e n t .

a Bacteremia

E.e.n 31 i m Bactereiia - Eglin

E.coli 3 i i m - Uli» 3J5 wA, U

F i g . 3: Mean plasma l e v e l s o f a n t i t h r o m b i n I I I i n s e p t i c p i g s w i t h o u t (a) and w i t h i n h i b i t o r (b) t r e a t m e n t .

58

Conclusions

Treatment of e x p e r i m e n t a l s e p t i c e m i a i n pi g s e x c l u s i v e l y by e g l i n r e v e a l e d a c o n v i n c i n g p r o t e c t i v e e f f e c t of t h i s p r o t e i n ­ase i n h i b i t o r on lun g t i s s u e and consumption o f plasma p r o t e i n s probably due t o i n h i b i t i o n o f u n s p e c i f i c p r o t e o l y s i s . Since, however, some severe organ f a i l u r e s (e.g. r e n a l i n s u f f i c i e n c y ) due t o septicemia c o u l d not be overcome, a c o m b i n a t i o n o f the i n h i b i t o r a d m i n i s t r a t i o n w i t h an approved i n t e n s i v e care t h e r a ­py (e.g. catecholamines, r e s p i r a t o r y t h e r a p y ) i s needed f o r an o p t i m a l outcome.

Acknowledgement

We are very g r a t e f u l t o Drs. H.-P. S c h n e b l i , W. Märkl and M. L i e r s c h (Fa. Ciba Geigy, Basel, S w i t z e r l a n d ) f o r s u p p l y i n g us w i t h cloned E g l i n c (^-N 0^-Acetyl-Eglin c, CGP 32968) as w e l l as t o Dr. R. Maschler and Prof. Dr. Dr. E. Fink (Fa. P l a n t o r g a n , Bad Zwischenahn, FRG) f o r E g l i n b from Hirudo m e d i c i n a i i s . We a l s o thank Drs. U. Seemüller and W. Müller-Esterl f o r making a v a i l a b l e anti-serum t o E g l i n c.

59

References

1 F r i t z , H. , M. Jochum, K.-H. Duswald, H. D i t t m e r , H. K o r t -mann, S. Neumann, H. Lang. 1984. Granulocyte p r o t e i n a s e s as m e d i a t o r s of u n s p e c i f i c p r o t e o l y s i s i n i n f l a m m a t i o n . A r e ­view. I n : Selected Topics i n C l i n i c a l Enzymology, V o l . 2 (eds.: D. M. Goldberg, M. Werner). Walter de G r u y t e r , Ber­lin-New York, 306-328.

2 Jochum, M., J. W i t t e , H. S c h i e s s l e r , H. K. Selbmann, G. Ruckdeschl, H. F r i t z . 1981. C l o t t i n g and o t h e r plasma f a c ­t o r s i n e x p e r i m e n t a l endotoxemia: i n h i b i t i o n o f d e g r a d a t i o n by exogenous p r o t e i n a s e i n h i b i t o r s . Eur. surg. Res. 13, 152-1 68.

3 Seemüller, U., M. Meier, K. Ohlsson, H. P. Müller, H. F r i t z . 1977. I s o l a t i o n and c h a r a c t e r i z a t i o n of a low molecular weight i n h i b i t o r ( o f chymotrypsin and human g r a n u l o y t i c e l a s t a s e and c a t h e p s i n G) from leeches. Hoppe-Seyler 1s Z. P h y s i o l . Chem. 358, 1105-1117.

4 Rink, H., M. L i e r s c h , P. Sieber, F. Meyer. 1984. A l a r g e fragment approach t o DNA s y n t h e s i s : t o t a l s y n t h e s i s of a gene f o r the protease i n h i b i t o r e g l i n c from the l e e c h Hirudo m e d i c i n a l i s and i t s expression i n E . c o l i . N u c l e i c Acids Res. 12, 6369-6387.

5 W e l t e r , H. F., 0. T h e t t e r , M. Siebeck, H. Wiesinger, U. E l ­s t e r , M. Jochum. 1985. Study on the t r e a t m e n t of shock lu n g w i t h superoxide dismutase (SOD) and C 1 - i n a c t i v a t o r (C1-INA). I n : Langenbecks Arch., Suppl. C h i r . Forum (ed.: F. S t e l z ­n e r ) . S p r i n g e r - V e r l a g B e r l i n - H e i d e l b e r g , 63-67.

6 Lewis, F. R., V. B. E l i n g s , J.A. Sturm. 1979. Bedside meas­urement of lung water. J. Surg. Res. 27, 250-261.

7 M a n c i n i , G., A. 0. Carbonara, J. F. Heremans. 1965. Immuno­chemical q u a n t i t a t i o n of a n t i g e n s by s i n g l e r a d i a l immuno­d i f f u s i o n . Immunochemistry 2, 235-254.