Table Of ContentLucio Miele · Spyros Artavanis-Tsakonas
Editors
Targeting
Notch in
Cancer
From the Fruit Fly to the Clinic
Targeting Notch in Cancer
Lucio Miele • Spyros Artavanis-Tsakonas
Editors
Targeting Notch in Cancer
From the Fruit Fly to the Clinic
Editors
Lucio Miele Spyros Artavanis-Tsakonas
Department of Genetics Department of Cell Biology
Louisiana State University Health Harvard Medical School
Sciences Center Boston, MA, USA
New Orleans, LA, USA
ISBN 978-1-4939-8857-0 ISBN 978-1-4939-8859-4 (eBook)
https://doi.org/10.1007/978-1-4939-8859-4
Library of Congress Control Number: 2018957844
© Springer Science+Business Media, LLC, part of Springer Nature 2018
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Preface
In 1991, a rare chromosomal translocation affecting chromosomes 9 and 7 was
described in T-cell lymphoblastic leukemia. The translocation involved TAN-1, a
previously unknown human locus highly homologous to Drosophila Notch, a gene
well known to developmental biologists as a determinant of cell fate during devel-
opment. This serendipitously discovered link between developmental biology and
cancer biology touched off a veritable explosion of discoveries on the role of Notch
in human malignancy. Today, major pharmaceutical and biotechnology companies
have Notch programs and have developed investigational drugs and biologics tar-
geting Notch signaling. Notch ligands have been used successfully to expand human
cord blood progenitor cells for transplantation purposes. Studies of Notch signaling
and its crosstalk with other developmental pathways have revealed a remarkably
complex network of molecular interactions at the core of the cell fate-controlling
machinery. Interest in this field has steadily increased, and today Notch signaling is
known to play roles in virtually every aspect of cancer biology, from control of dif-
ferentiation, proliferation, and apoptosis in transformed cells to angiogenesis,
tumor-stroma interaction, and anticancer immune responses. A number of observa-
tions have revealed a role of Notch in the self-renewal of “cancer stem-like cells” or
“tumor-initiating cells” that are thought to be a major cause of treatment failure in
cancer. It is not unreasonable to speculate that pharmacological manipulation of
Notch signaling could alter the practice of medicine in the treatment of many human
malignancies. This does not mean that targeting Notch in the clinic will be easy or
straightforward. The notorious context dependence of Notch effects, well known to
developmental biologists but frustrating to pharmacologists and cancer biologists,
means that the question “what does Notch do in cancer and what is the best strategy
to target it” does not have a simple answer.
Advances in genomics have revealed that human cancers are remarkably plastic,
particularly during and after treatment with chemo- or radiotherapy or even targeted
therapy: many cancers undergo a process of quasi-Darwinian clonal evolution,
selecting cellular clones with more stem-like characteristics. Phenotypic plasticity
due to tumor microenvironmental effects can also alter the biology of cancer cells
v
vi Preface
through metabolic and/or epigenetic effects. These aberrant cell fate decisions
prominently involve developmental pathways including Notch.
Efforts to translate our growing but still incomplete understanding of Notch biol-
ogy in cancer will have to take these complexities into account. A detailed under-
standing of the intricate crosstalk between Notch and other pathways of therapeutic
interest will be necessary to design rational drug combinations for specific diseases
and disease subsets. The multiple classes of agents currently in various stages of
development have different advantages and disadvantages. It is still unclear what
the best agents are or what drug combinations are most promising in individual
indications. Determining the status of Notch signaling in tumor samples is a chal-
lenge in its own right, with four Notch paralogs, five ligands, a host of co-ligands,
as well as canonical and noncanonical Notch target genes that are differentially
affected in different cell types. These hurdles are not insurmountable, but they need
to be considered by those approaching the clinical development of Notch-targeting
agents if we are to avoid unpleasant surprises.
Genetic experiments in model organisms have revealed much about genes and
pathways that modify the effects of Notch signaling in tumorigenesis. These invalu-
able insights are now being translated to human tumor biology and experimental
oncology.
This volume is an attempt at describing the current state of the art in the field of
Notch signaling in cancer, with a specific focus on targeting Notch signaling for
therapeutic purposes. Internationally known experts in the Notch field have contrib-
uted chapters to what we hope will be a comprehensive discussion.
No single book can encompass all aspects of a vast and growing field of biomedi-
cal research, and this volume is no exception. We hope that the reader will be left
with a clear view of the field’s complexity, a clear understanding of what is known,
and a sense of what remains unknown. We thank the readers for their interest in our
work and hope that our effort will stimulate further interest in this fascinating field
of biomedical research.
New Orleans, LA, USA Lucio Miele
Boston, MA, USA Spyros Artavanis-Tsakonas
Contents
1 Structural Biology of Notch Signaling . . . . . . . . . . . . . . . . . . . . . . . . . 1
Kelly L. Arnett, Tom C. M. Seegar, and Stephen C. Blacklow
2 Noncanonical Notch Signaling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Jyothi Vijayaraghavan and Barbara A. Osborne
3 Dual Function of Notch Signaling in Cancer:
Oncogene and Tumor Suppressor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Ute Koch and Freddy Radtke
4 Out on the Fringe: Modulation of Notch Signaling
by Glycosylation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
Keli Xu and Sean E. Egan
5 Notch Signaling: A Pivot Regulator of Adaptive
and Innate Immunity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
Takumi Kumai and Paulo C. Rodriguez
6 Notch in Ovarian Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153
Emily Gerry, Vivek Singh, and Tian-Li Wang
7 Notch Signaling in Graft-Versus-Host Disease . . . . . . . . . . . . . . . . . . . 175
Lisa M. Minter
8 Notch Signaling in T-Cell Acute Lymphoblastic
Leukemia and Other Hematologic Malignancies . . . . . . . . . . . . . . . . . 199
Catherine Hoofd, Vincenzo Giambra, and Andrew P. Weng
9 The Role of Notch in Breast Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
Jeffrey C. Bloodworth and Clodia Osipo
10 Notch in Lung Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241
Sara L. Sinicropi-Yao, Michael J. Koenig, and David P. Carbone
vii
viii Contents
11 Notch Signaling in Pediatric Soft Tissue Sarcoma . . . . . . . . . . . . . . . . 277
Cristina Cossetti, Alberto Gualtieri, Silvia Pomella,
Elena Carcarino, and Rossella Rota
12 Notch Ligands in Hematopoietic Stem Cell Production . . . . . . . . . . . 313
Anna Bigas, Cristina Ruiz-Herguido, Rosa Aligué,
and Lluís Espinosa
13 Notch Signaling in the Normal Intestine and Intestinal Cancer. . . . . 333
Lluís Espinosa, Erika López-Arribillaga, Oriol Bachs,
and Anna Bigas
14 N otch Signaling in Estrogen-Dependent Cancers . . . . . . . . . . . . . . . . 353
Judy S. Crabtree
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 381
Chapter 1
Structural Biology of Notch Signaling
Kelly L. Arnett, Tom C. M. Seegar, and Stephen C. Blacklow
Abstract The conserved Notch signaling pathway plays a central role in devel-
opment and adult tissue homeostasis. Notch signaling is initiated by binding to a
transmembrane ligand. E3 ubiquitin ligase-mediated ligand endocytosis enables
release of the negative regulatory region (NRR) of Notch from autoinhibition,
which then allows metalloprotease cleavage within the NRR, followed by intra-
membrane cleavage by the γ-secretase complex. After release from the mem-
brane, the Notch intracellular domain translocates to the nucleus to form a
transcriptionally active complex and initiate transcription of Notch-responsive
genes. Structural studies of Notch and Notch-associated molecules, which have
advanced our understanding of each of these steps in the Notch signaling path-
way, are reviewed here.
Keywords Notch · DSL · Receptor signaling · Protein biochemistry · Structural
biology · Regulated intramembrane proteolysis · Transcription
1.1 Introduction
Notch receptors bind transmembrane ligands on neighboring cells and transduce
signals, playing an essential role in cell-fate decisions during development and
tissue homeostasis. In mouse, fly, and worm models, severe deficiencies in Notch
signaling lead to embryonic lethality. Abnormal decreases and increases in Notch
K. L. Arnett · T. C. M. Seegar · S. C. Blacklow (*)
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School,
Boston, MA, USA
Department of Cancer Biology, Dana Farber Cancer Institute, Boston, MA, USA
e-mail: [email protected]; [email protected];
[email protected]
© Springer Science+Business Media, LLC, part of Springer Nature 2018 1
L. Miele, S. Artavanis-Tsakonas (eds.), Targeting Notch in Cancer,
https://doi.org/10.1007/978-1-4939-8859-4_1
2 K. L. Arnett et al.
Ligand binding
Sending cell Receiving cell
and Receptor maturation:
Receptor activation Fringe glycosylation
and other and furincleavage
g-secretase:
glycosyltransferases
PS1, Nicastrin, Furin
Pen2, Aph-1
pro-Notch
Golgi
NECD Pofut
Mind bomb 1 NICD
Rumi ER
DSL
Ligand Ligand
endocytosis Adam10/17 Transcriptional
NICD activation
MAM target
CSL genes
CBP/p300
Co-repressor
CSL
Fig. 1.1 Overview of major events in Notch signaling. Notch is expressed as a pro-protein precur-
sor and undergoes a series of posttranslational modifications during maturation, including O-linked
glycosylation and processing at site S1 by a furin-like protease. Signals are initiated by the engage-
ment of ligand on the signal-sending cell with the extracellular part of Notch (NECD) on the sig-
nal-receiving cell. Ligand endocytosis promotes relief of Notch autoinhibition allowing
metalloprotease cleavage by an ADAM family metalloprotease at site S2. This proteolytic step
allows the cleavage of Notch by the γ-secretase complex at site S3 within the transmembrane
domain and release of the Notch intracellular domain (NICD) from the membrane. Upon
translocation to the nucleus, NICD enters into a transcriptional activation complex with the DNA-
binding transcription factor CSL and coactivator Mastermind (MAM)
signaling are associated with human developmental abnormalities and disease, and
cancer-associated mutations have been found which produce both constitutively
active and inactive forms of Notch. Notch receptors, their ligands, and many
accessory proteins are essential for transducing a Notch signal, which proceeds
through a series of connected events: (1) ligand binding, (2) ligand endocytosis, (3)
release from autoinhibition, (4) metalloprotease cleavage, (5) intramembrane cleav-
age, and (6) transcriptional activation (Fig. 1.1).
Notch receptors are large modular transmembrane proteins with distinct regions
playing specific roles in each of these steps (Fig. 1.2a). The N-terminal region of
Notch receptors consists of a series of epidermal growth factor (EGF)-like repeats
that bind ligands of the Delta and Serrate families and initiate the Notch signal [28,
94]. C-terminal to the EGF repeats, a region known as the negative regulatory region
(NRR) of Notch holds Notch receptors in an autoinhibited conformation prior to
ligand engagement [35, 95]. Ligand binding and endocytosis of the ligand, stimu-
lated by Mind bomb or Neuralized E3 ubiquitin ligases, are required for release
