Molecular Biology of the Arterial Wall

175 Pages·1987·10.45 MB·English

by  M. A. Lehrman

Veroffentlichungen aus der
Geomedizinischen Forschungsstelle
(Leiter: Professor Dr. Dres. h. c. G. Schettler)
der Heidelberger Akademie der Wissenschaften
Supplement zu den Sitzungsberichten der
Mathematisch-naturwissenschaftlichen Klasse
J ahrgang 1987
G. Schettler (Ed.)
Molecular Biology
of the Arterial Wall
Springer-Verlag
Berlin Heidelberg New York
London Paris Tokyo
Prof. Dr. Ores. h. c. Ootthard Schettler
Prasident der Heidelberger Akademie der Wissenschaften
Karlstrasse 4, 6900 Heidelberg, FRO
ISBN-13:978-3-540-17899-6 e-ISBN-13:978-3-642-83118-8
DOl: 10.1007/978-3-642-83118-8
Library of Congress Cataloging-in-Publication Data
Molecular biology of the arterial wall.
(VerOffentlichungen aus der Geomedizinischen Forschungsstelle der Heidelberger
Akademie der Wissenschaften) (Supplement zu den Sitzungsberichten der
Mathematisch-Naturwissenschaftlichen KJasse; Jahrg.)
Based on a conference held in Heidelberg, Germany, Sept_ 27 -Oct. 4, 1986 and
organized by the Heidelberg Academy of Sciences. I. Arteriosclerosis
Etiology-Congresses. 2. Arteries-Congresses. 3. Pathology, Molecular-Con
gresses. 4. Molecular biology-Congresses. I. Schettler, Gotthard. II. Heidelber
ger Akademie der Wissenschaften. III. Series. IV. Series: Supplement zu den
Sitzungsberichten der Mathematisch-Naturwissenschaftlichen Klasse; Jahrg. 1987.
(DNLM: 1. Arteries-congresses. 2. Molecular Biology-congresses. WG 510
M718)
RC692.M6 1987 612'.133 87-12843
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Table of Contents
Opening Address .....•........................... o. ... IX
0.0 •• 0.0. 0... •• •• •• ••
Workshop I
lipoproteIns and Lipoprotein Receptors
0 ••• 0 0 •••••••••••••• 0 •••••••• , •• 0 0 0 0 ••
LDL Receptor Mutations in Patients with Familial Hypercholesterolemia
M.A. Lehrman, H.H. Hobbs, M.S. Brown, J.L. Goldstein, DoW. Russel ..... 2
Lipoprotein Receptors and Their Ligands: Structure and Function of
Apoproteins E and B
R.W. Mahley ... ,., .................................................... ,. 5
The Role of HDL in Cholesterol Homeostasis of Macrophages and Reversed
Cholesterol Transport
G. Schmitz, H. Robenek .................................... ,............ 8
Lipoproteins, Macrophages and Atherosclerosis
0.......
D. Steinberg ........ , ................... , ....................... 10
Arterial Lipoprotein Metabolism: Site-Specific Differences Related to
Susceptibi Iity to Atherosclerotic Lesions
T.E. Carew, D.C. Schwenke ............................................. 13
Lipoprotein Disposition by the Liver: A Concerted Action of Various
Cell Types
T.J.C. van Berkel, L. Harkes, J. F. Nagelkerke, J. K. Kruijt.............. 17
The Macrophage and Endothelial Cell Scavenger Receptor
D.P. Via, A. Fanslow, W.E. Koff, H.A. Dresel. ........................... 19
Characterization of the Hepatic Scavenger Receptor
H.A. Dresel, E. Friedrich, H.J. Sinn, G. Schettler, D.P. Via ......... 22
0 ••••
Workshop II
Growth Factors .......................................... 25
0 ••••••••••••• 0 • • ••
The Role of PDGF in Health and Disease
R. Ross .......................................... 26
0 ••••••••••••••••••••••
Role of PDGF-Iike Growth Factors in Autocrine Stimulation of Cell Growth
C .-H. Heldin ............................•.....•.......... 30
0 •••••• 0 •••• •••
The PDGF Receptor as a Mediator of Mitogenesis
LoT, Williams, J .A. Escobedo, M.T. Keating, T ,0. Daniel, S.R. Coughlin .. 32
VI
Alpha- and Beta-Heparin Binding Growth Factors: Chemistry and Biology of
Acidic and Basic Fibroblast Growth Factors
A. Baird, P. Cuevas, N. Ueno, F. Esch, N. Ling ............•.............. 36
Structure, Homologies and Activities of Acidic Fibroblast Growth Factor
K.A. Thomas, G. Gimenez-Gallego, J. DiSalvo, D. Linemeyer, L. Kelly,
J. Menke................................................................ 41
Endothelial Cell Growth Factor and its Receptor
T. Maciag, W.H. Burgess ................................., ............... 44
Workshop III
Endocytosis and Exocytosis .................................................. 47
Intracellular Transport of Human Class I Antigens and an Adenoviral
Glycoprotein
P.A. Peterson, M. Andersson, I. Martens, S. Piiiibo, L. Severinsson .......• 48
Molecular Mechanisms of Endocytic Sorting
I. Mellman, S. Schmid, R. Fuchs, M. Marsh, V. Lewis, T. Koch, S. Green,
H. Plutner, A. Helenius. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 51
Characterisation of a Fusion Event from the Endocytic Pathway
J. Davey, G. Warren. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . • . . . . . .. 56
Membrane Boundaries Involved in the Selective Trafficking of Cell Surface
Receptors
C.R. Hopkins ............................................................ 59
The Molecular Basis of Human Cellular Iron Metabolism
R.D. Klausner, J.B. Harford ..........................•..........•....... 61
Workshop IV
ONC Genes and Proliferative Disease........................................ 65
Molecular Features Involved in Cell Surface Receptor Function and Their Role
in Oncogenesis
H. Riedel, Y. Yarden, A. Gray, L. Coussens, T. Dull, J. Sch lessinger,
A. Ullrich ........................... " . .. . . . . . . . . . . . . . . .. . .. . . . . . . . . . . . 66
Transforming Activity of the fms Oncogene (CSF-1 Receptor)
C.J. Sherr, M.F. Roussel~F. Wheeler, C.W. Rettenmier.... .•... ....... 69
Functional and Structural Analysis of the v-src and c-src Genes
H. Hanafusa ...........................~ ......~ . . . . . . . . . • . . . • • • . . . 71
Ras Oncogenes and Cell Transformation
C.J. Marshall ................................................... · .... ··. 73
Cis Regulatory Control of mos Oncogene Expression
-M.L. McGeady, T.G. Wood, D.G. Blair, A. Seth, F. Propst, M. Oskarsson,
M. Schmidt, G. Vande Woude. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
Analysis of the Function of the fos Oncogene Product
R. Muller, T. Jenuwein, M.Renz ........................................ 81
VII
Cooperative Interactions Involving Cellular and Viral Oncogenes in the
Development of Malignant Tumors
E. Mougneau, C. Cerni, F. Cuzin ......................................... 84
Workshop V
White Blood Cells........................................................... 85
The Motor of Leukocytes and Platelets: 1986
T.P. Stossel, J.H. Hartwig, P.A. Janmey, H.L. Yin, K.S. Zaner ........... 86
Heterogeneity of Murine and Human Fcy Receptors
J . C. Unkeless........................................................... 88
Complement Receptors
D.T. Fearon............................................................ 91
Cachectin: A Macrophage Protein that Induces a Catabolic State in Infected
Animals
A. Cerami. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 94
The Role of Cytosolic Free Calcium, (Ca2+)., and Phosphoinositides as
Intracellu lar Signals I
P.O. Lew •............................................................... 95
The Macrophage-Blood Interface: Cell Localisation and Recruitment in
Bone Marrow, Liver and Brain
S. Gordon, P.R. Crocker, L. Morris, H.V. Perry ......................... 97
Normal and Neoplastic Early Lymphocyte Maturation
I. Weissman, C. Muller, G. Tidmarsh, C. Okada, C. Whitlock .............. 101
Molecular Regulation of Complement Gene Expression
H.R. Colten ......................................................•...... 102
Workshop VI
Prostaglandins and Leukotrienes ......................................•..... 105
The Role of GTP-Binding Proteins in Coupling Prostaglandin and Leukotriene
Receptors to Intracellular Second Messenger Systems
P. Gierschik, K.H. Jakobs ............................................... 106
Regulation of Vascular Tone by Prostaglandins and Endothelium-Derived
Relaxing Factor
U. Forstermann......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . • . • • . . . . . • . . . . .. 110
Endogenous Cysteinyl Leukotrienes as Mediators in Endotoxin Shock and
Tissue Trauma
D. Keppler, W. Hagmann, C. Denzl inger, S. Rapp, M. Huber, A. Guhlmann,
S. Kastner.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . • • . . . . . . .. 115
Source and Mechanism of Release of Arachidonic Acid in Blood Platelets
J.B. Smith .............................................•.....••..•••.... 117
Eicosanoids and the Pulmonary Circulation
W. Seeger, N. Suttorp, H. Neuhof ........................................ 119
VIII
Regulation of the Production (Leukotriene All Hydrolase) and the Action
(L~~:~r~~~~~t!fu'!:~:s~~.~I.t~~~ .~~ ~~~.~~~~~~~.~~
.......•....•.•.•..••....... 123
Enzymic Synthesis and Degradation of Prostaglandin D2
O. Hayalshi, Y. Urade, K. Watanabe ...................................... 126
Receptor-Dependent Regulation of Human Polymorphonuclear Leukocyte
Responses to Leukotrienes .
C.H. Koo, L. Baud, T. Marotti, M. Cheung, J.P. Harvey, E.J. Goetzl ...... 129
Leukotriene C -Binding Proteins
K.F.Auste~ ..........................•.................••........•..... 132
Leukotriene C Metabolizing Enzymes
L. tlrning, K. Bernstrom, M. SOderstrom, B. Mannervik,
S. Hammarstrom......................................................... 134
Workshop VII
Biology of Smooth Muscle and Endothelium ................................... 139
The Fibrinolytic System of Cultured Bovine Aortic Endothelial Cells
D.J. Loskutoff.......................................................... 140
Tumor Necrosis Factor /Cachectin and the Modulation of Endothelial Cell
Coagulant Properties
D.M. Stern, P.P. Nawroth ...........................•...........•.•..... :145
Membrane Glycoproteins of Endothelial Cells and Platelets
J.A. van Mourik, J.C. Giltay, O.C. Leeksma, J. Zandbergen-Spaargaren .. 148
The Molecular Biology of Endothelial Cell von Willebrand Factor
D.C. Lynch ......................................................•...•.. 151
Induction and Regulation of Endothelial Gene Expression
T. Collins, J.S. Pober ................................................... 155
Endothelium-Vascular Smooth Muscle Interactions in Culture
R.W. Alexander, J. Leopold, K. Griendling, P. Ganz ...................... 159
Endothelial Morphogenesis
S.M. Schwartz, R.L. Heimark .•.................................•....•.•. 163
Closing Remarks
S.C. Silverstein ................................•..•....•.•.•.••.••.••... 166
Final Word
G. Schettler ..................................•.....••...••..•.....•.•... 167
List of Participants. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . • . . . . . . • . . . . . . . • • . . • • . . • . .. 168
List of Sponsors ...........................•..•......•..••.••...•......•••.. 169
OPENING ADDRESS
by G. Schettler
This conference on Molecular Biology of the Arterial Wall has been organized by
the Heidelberg Academy of Sciences and a group of truly outstanding international
experts who are acting as chairpersons. It is intended to be the first in a series of
scientific meetings organized by the Academy in the years to come focusing on
special topics related to basic medical research. Backed by a prestigious tradition.
the Academy seeks to create an open forum for discussions of wide-ranging develop
ments in basic research and for interdisciplinary scientific cooperation among Inter
national scientists of world renown.
More and more frequently in the last decades. researchers guided by quite different
basic concepts and using different methods have obtained results which can be
synthesized in unexpected ways to produce interesting conclusions that require us
to view scientific questions in a new light. After a long period in which researchers
working on the biology and pathobiology of the arterial wall were somewhat isolated
from the mainstream of cellular biology. it seems that as cellular biologists approach
the level of single cells and single genes. we are also developing a more fundamental
understanding of the pathomechanisms of diseases of the arterial wall. The situation
in the past was often frustrating. Cellular interactions in the arterial wall were too
complex to be studied more specifically. Furthermore. arterial lesions evolve very
slowly. which seemed to hinder an experimental approach. Finally. there was the
attractive and popular lipid hypothesis that plasma components can be a sufficient
cause for atherosclerosis and its clinical syndromes. This hypothesis has encouraged
the study of atherogenesis in terms of plasma lipoproteins and apolipoproteins. which
are available in large quantities and can be screened for mutations .. However. dis
eases and cellular changes in the arterial wall are not just a consequence of hyper
lipidemia and an increased influx of lipids into the arterial wall. The atherosclerotic
plaque is not a degenerative process. As already hypothesized by the Cerman
pathologist Rudolf Virchow in 1856. specific injuries of the arterial wall cause a
disturbance of the critical balance between cell proliferation and cell destruction.
Following this assumption. one can postulate that specific lesions in the arterial wall
are constantly forming. regressing. or progressing.
As new cellular and biochemical techniques have become available in the last years.
researchers have developed new tools to address their investigations to the cellular
basis of both normal arterial homeostasis and the diseased state when lesions of the
arterial wall are developing. Consequently. our knowledge of the biology of arterial
wall cells and molecular interactions. e.g .• on the level of cellular receptors. is ex
panding more and more rapidly. It is becoming evident that many of the new notions
will contribute to generating a basis for an understanding of Virchow's hypothesis
on a molecular level.
Workshop I
Lipoproteins and Lipoprotein Receptors
LDL Receptor Mutations in Patients with Familial Hypercholesterolemia
M.A. Lehrman, H.H. Hobbs, M.S. Brown, J.L. Goldstein, and D.W. Russell
Department of Molecular Genetics, University of Texas Health Science Center at
Dallas, 5323 Harry Hines Blvd., Dallas, Texas 75235 USA
INTRODUCTION
The low-density lipoprotein (LDL) receptor plays a major role in the maintenance
of normal LDL level in humans (1). Mutations in the LDL receptor gene result
in Familial Hypercholesterolemia (FH), an autosomal dominant disorder charac
terized by elevated levels of plasma LDL (1). The LDL receptor is normally
synthesized in the endoplasmic reticulum (ER) as a precursor of 120,000 daltons
that contains immature N- and O-linked oligosaccharides. During synthesis, an
amino-terminal signal sequence is cleaved from receptor. Maturation of the
oligosaccharides in the Golgi complex generates the mature form of the receptor
with a molecular weight of 160,000. The receptors are then transported to the
cell surface, where they cluster in coated pits and can bind LDL. By receptor
mediated endocytosis, the bound LDL is internalized as the coated pits
invaginate to form coated vesicles. The receptor and ligand are then trans
ported to endosomes, where they dissociate. The LDL is eventually transported
to lysosomes where it is degraded. In contrast, the receptor returns to the
cell surface to repeat the cycle (1).
The LDL receptor gene consists of eighteen exons that span at least 40 kb qf
DNA. Exon 1 encodes a cleavable signal sequence. Exons 2 through 18 encode
the mature receptor protein, which consists of 839 amino acids and is thought
to be composed of five domains (1). Listed from the amino to the carboxy
terminus, they are: 1) a putative ligand binding domain of 292 amino acids
(exons 2-6 of the receptor gene); 2) a domain of 417 amino acids with homology
to the epidermal growth factor (EGF) precursor (exons 7-14); 3) a domain of 58
amino acids rich in serine and threonine-linked oligosaccharides (exon 15); 4)
a 22 amino acid membrane-spanning region (exons 16 and 17); and 5) a cyto
plasmic domain of 50 amino acids (exons 17 and 18). In order to begin to
understand the functional roles of these domains, we have characterized the
mutations that occur in patients with FH by a combination of DNA and RNA
blotting, SI nuclease mapping, molecular cloning, and DNA sequencing. These
studies have also provided insight into the genetic basis for these mutations
and the mechanisms by which the receptor gene can be rearranged.
RESULTS AND DISCUSSION
Two mutations have been identified that eliminate part of the putative ligand
binding domain. The mutation in one patient, FH 626, which was characterized
by cloning and sequencing, was caused by a 0.8 kb deletion that eliminated exon
5, and generates a receptor that is smaller than normal and does not bind LDL
(2). Thus, this domain of the receptor must function in the binding of ligand.
A second mutation, FH 563 (3) lost approximately 30 nucleotides (10 amino
acids) in exon 4, as judged by SI nuclease mapping. This receptor gets
transported to the cell surface at a very slow rate. Since assays that measure
LDL binding require that the receptors be present on the cell surface (1), it
is not known if this mutation in the binding domain affects the ability of the
receptor to bind LDL.