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Logo of nihpaAbout Author manuscriptsSubmit a manuscriptNIH Public Access; Author Manuscript; Accepted for publication in peer reviewed journal;
Clin Cancer Res. Author manuscript; available in PMC Jun 15, 2011.
Published in final edited form as:
PMCID: PMC3008160

Targeting Cancer Stem Cells through Notch Signaling


The cellular heterogeneity of neoplasms has been at the center of considerable interest since the “cancer stem cell hypothesis”, originally formulated for hematologic malignancies, was extended to solid tumors. The origins of cancer “stem” cells (CSCs) or tumor-initiating cells (TICs) (henceforth referred to as CSCs) and the methods to identify them are hotly debated topics. Nevertheless, the existence of sub-populations of tumor cells with stem-like characteristics has significant therapeutic implications. The stem-like phenotype includes indefinite self-replication, pluripotency and, importantly, resistance to chemotherapeutics. Thus, it is plausible that CSC, regardless of their origin, may escape standard therapies and cause disease recurrences and/or metastasis after apparently complete remissions. Consequently, the idea of selectively targeting CSCs with novel therapeutics is gaining considerable interest. The Notch pathway is one of the most intensively studied putative therapeutic targets in CSC, and several investigational Notch inhibitors are being developed. However, successful targeting of Notch signaling in CSC will require a thorough understanding of Notch regulation and the context-dependent interactions between Notch and other therapeutically relevant pathways. Understanding these interactions will increase our ability to design rational combination regimens that are more likely to prove safe and effective. Additionally, to determine which patients are most likely to benefit from treatment with Notch-targeting therapeutics, reliable biomarkers to measure pathway activity in CSC from specific tumors will have to be identified and validated. This article summarizes the most recent developments in the field of Notch-targeted cancer therapeutics, with emphasis on CSC.

Cancer “stem” cells and treatment resistance

Despite decades of search for an elusive “magic bullet”, the pharmacological treatment of cancer still relies heavily on traditional chemotherapy, which is being slowly supplemented by targeted agents. Incremental improvements are being made, but treatment resistance remains a major cause of morbidity and mortality.

For patients, clinicians and cancer biologists, the most frustrating feature of malignancies is their inherent adaptability. In the clinic, this adaptability translates into drug-resistant disease recurrence and metastasis, often after clinical and even pathological complete responses.

One possible explanation for the biological plasticity of cancers is their cellular heterogeneity. In recent years, a distinct cellular hierarchy has been identified in several hematopoietic and solid tumors. Many cancers appear to contain a small population of pluripotent “tumor initiating cells” or “cancer stem cells” (CSC) [1-14]. The CSC hypothesis states that CSC possess some of the biological properties of normal stem cells, including indefinite self-replication, asymmetric cell division, and resistance to toxic agents, due in part to elevated expression of ABC transporters. Normal tissue stem cells are characterized by very slow proliferation rates and this is generally assumed to be the case for CSC. However, it is not established that all cancer cells with stem-like markers always proliferate slowly in vivo. Additionally, CSC proliferation rates may depend on microenvironment, type of oncogenic mutations, stage of malignancy and other variables. The origin of CSC is the subject of considerable debate. One popular version of the “CSC hypothesis” proposes that CSC originate from the transformation of normal tissue stem cells, and give rise to other cancer cells through a process of aberrant differentiation that resembles that of normal tissues but evades physiological regulatory mechanisms [11]. CSCs from various primary tumors or cell lines do express tissue stem cells markers, including CD133 (promimin-1), nestin, c-kit, sox2, Oct4, and musashi-1 (Figure 1A) [5, 10, 15-18]. On the other hand, it is also conceivable that “stemness” is a phenotype that can be acquired through transformation or modulated by extracellular signals. Stemness can be restored in normal fibroblasts by expression of Oct4, Sox2, Nanog, and LIN28 [19], or Oct3/4, sox2, Klf4, and c-Myc [20]. Physiological de-differentiation has been described in vertebrate and invertebrate non-neoplastic systems [21]. For example, spermatogonia in the Drosophila testis can restore male germline stem cells by a JAK-STAT-dependent de-differentiation process [22, 23]. A well-known process of partial de-differentiation in epithelial cancers is epithelial-mesenchymal transition (EMT) [24]. This consists of loss of epithelial-specific cytokeratins and E-cadherin, and acquisition of mesenchymal markers such as vimentin and N-cadherin. Transcription factors such as Twist, Snail or Slug and TGF-β family secretory factors can induce EMT. A model supported by recent evidence from the Weinberg group [25] is that “stemness” can be re-acquired by cancer cells when they undergo EMT, a process associated with an invasive and metastatic phenotype. An intermediate possibility between the notion that CSC derive exclusively from normal tissue stem cells and the notion that stemness can be acquired by any cell is the suggestion that CSCs may originate from few cell populations, including tissue stem cells and immature progenitors capable of short-term self-replication. Dontu et al. [26] suggested that estrogen receptor (ERα)-negative breast cancers and poor-prognosis ERα-positive cancers (presumably Luminal B) arise from ERα-negative primitive mammary stem cells, while less aggressive ERα-positive cancers (presumably luminal A) arise from ERα-positive intermediate progenitors.

Figure 1
A: schematic representation of an idealized CSC. The figure shows a list, not meant to be all-inclusive, of pathways that modulate the CSC phenotype. CSC exist in the context of “niches” formed by neighboring cells and extracellular matrix ...

These models, schematically represented in Figure 1B, may not be mutually exclusive and may apply to different malignancies, different subtypes or stages of the same malignancy. A common element in the different versions of the CSC hypothesis is the concept of a cellular hierarchy in solid tumors and hematological malignancies cancers similar to that of normal tissues. CSC are thought to be capable of asymmetric cell division that maintains the CSC population and produces pluripotent “progenitor-like” cells. These in turn give rise to the “bulk” tumor cells through proliferation and aberrant “differentiation” (Figure 1C). Because of their ability to regenerate all cell types in a tumor, CSC, and possibly progenitors, are thought to have higher tumorigenic potential than “bulk” tumor cells.

A factor that complicates the testing of this model in human tumors is the fact that the identification of CSC as opposed to “bulk” tumor cells in clinical specimens relies on markers that are different for different malignancies and are not universally accepted [2, 27]. A widely used functional test involves the measurement of limiting dilution tumorigencity in immunodeficient NOD/SCID mice, with CSC being highly tumorigenic in very small numbers, unlike “bulk” tumor cells. However, the tumorigenicity of human cancer cells in mice depends heavily on the degree of immunodeficiency. When melanoma cells were injected into more permissive NOD/SCID IL-2 receptor γ chain knockout mice (“NOG” mice), that lack NK cells, the fraction of tumorigenic cells appeared be in the order of 25-27%, inconsistent with the rarity of tumorigenic cells identified in NOD/SCID models [28, 29]. These observations indicate that the role of immune surveillance, both innate and adaptive, in defining CSC requires careful investigation, and have led some investigators to question the CSC hypothesis. In addition to the immune system, an additional complicating factor is the role of cellular microenvironment, or CSC “niches”, where CSCs survive inside the primary tumor or at distant sites [30]. Particularly in view of the fact that the stem-like phenotype may be inducible by paracrine signals such as TGF-β, Wnt and Hedgehog and signals transmitted by cell-cell contact such as Notch, the importance of microenvironment in determining the biological properties of CSCs should not be underestimated. That said, clinical evidence supporting the existence of CSCs and their role in treatment resistance has emerged, particularly in breast cancer. Li et al. showed that tumorigenic cells with stem-like markers are selected by neo-adjuvant chemotherapy [31]. Creighton et al. have recently shown that breast cancer cells surviving in patients after treatment with either docetaxel or letrozole have gene expression signatures characteristic of stem-like and EMT phenotypes [32]. Taken together, the evidence available today suggests that cells with a stem-like phenotype are found in several human malignancies, and that, to the extent that they exist according to current hypotheses, these cells are not adequately targeted by currently used cancer therapeutics. It is reasonable to hypothesize that a complete eradication of these cells will be necessary to attain long-lasting remissions or cures. A strategy that is receiving considerable attention is to target CSC through evolutionarily ancient pathways that control self-renewal and cell fate decisions in undifferentiated, pluripotent cells [27]. Many such pathways have been identified (Figure 1A). Most of these are evolutionarily conserved and have multiple developmental roles, as well as intricate cross-talk interactions. Among them, Wnt [33], Hedgehog [34], and Notch [14, 35] are the focus of intense developmental therapeutics efforts. This article will focus on Notch signaling as a putative therapeutic target in CSC.

Notch signaling: mechanistic complexity with potential therapeutic implications

The Notch pathway is a short-range communication system in which contact between a cell expressing a membrane-associated ligand and a cell expressing a transmembrane receptor sends the receptor-expressing cell (and possibly both cells) a cell fate regulatory signal. This signal takes the form of a cascade of transcriptional regulatory events that affects the expression of hundreds of genes, and has profound, context-dependent phenotypic consequences. Several recent articles discuss the biochemical features of the pathway [36-38] and its possible roles in cancer [39-49].

Mature Notch receptors (in mammals Notch-1 through -4) are non-covalent heterodimers consisting of an extracellular subunit (NEC), and a transmembrane subunit (NTM, Figure 2). NEC contains multiple EGF-like repeats and three specialized Lin-Notch repeats (LNR) that form a tight hydrophobic interaction with the extracellular stump of NTM, masking an ADAM (A Disintegrin and Metalloprotease) cleavage site. The region of interaction between the two subunits is called HD (Heterodimerization Domain). Canonical Notch ligands are also transmembrane proteins (Figure 2) with multiple EGF-like repeats, a short cytoplasmic tail and a specialized DSL (Delta-Serrate-Lag2) domain at the N-terminus. Ligand binding triggers dissociation of NEC from NTM, unmasking the ADAM cleavage site (Figure 3A, B). NEC is trans-endocytosed into ligand-expressing cells while NTM is cleaved at the membrane by an ADAM, generating an intermediate called NEXT (Notch Extracellular Truncation). The latter is further cleaved by γ-secretase, generating an active fragment (NIC) (Notch Intracellular) or NICD (Notch Intracellular Domain, Figure 3A). NIC is transported into the nucleus (Figure 3D) where it binds ubiquitous transcription factor CSL, (CBF-1, Suppressor of Hairless, Lag-1), also known as RBP-jκ in mice. NIC binds to CSL and displaces a large co-repressor complex containing SKIP, SHARP, histone deacetylases and other co-repressors. The NIC-CSL complex recruits a co-activator complex containing a Mastermind-like protein (MAML1-3 in mammals) as well as p300 and other chromatin-modifying enzymes, forming the Notch Transcriptional Complex (NTC), and activating transcription [38]. CSL target genes are numerous and include HLH-family negative transcriptional regulators of the HES and HEY family, but also oncogenes such as c-Myc. In the nucleus, NIC is phosphorylated by CDK8, ubiquitinated by Sel10/FWB7 and degraded by the proteasome, thus terminating the signal. Putative non-canonical pathways have been suggested but remain incompletely characterized. Among them, physical interaction of Notch-1IC with the IKK signalosome [50], with nuclear IKKα [51, 52] and with p50 [53, 54] and of Notch-3IC with cytoplasmic IKKα [55] may mediate therapeutically relevant cross-talk with NF-κB. Physical interaction of Notch-1IC with p85 PI3-kinase α [56] may mediate non-nuclear cross-talk with AKT, leading to survival signaling [57].

Figure 2
Schematic structure of Notch receptors and ligands
Figure 3
Diagram of Notch activation and canonical signaling mechanisms

A closer look at this apparently simple signaling pathway reveals a dizzyingly intricate series of mechanisms that finely regulate the timing, intensity and biological consequences of Notch signaling, and are likely to have significant therapeutic implications. The main factors that should be taken into consideration when Notch signaling in the clinical seeting are listed below:

1) Paralog-specific effects

Mammals have four Notch paralogs, which differ mostly in the number of EGF repeats and the C-terminal part of NIC. Although in theory they all signal through the same mechanisms, in many systems they have non-overlapping and even opposing effects [58-61]. This is relevant when comparing agents that block all Notch signaling to agents that selectively block a single receptor. In our hands, siRNA knockdown of different Notch receptors in breast cancer cells affects the expression of different gene sets (Pannuti et al., in preparation). This is consistent with data indicating that Notch-2, unlike Notch-1, -3 and -4, may have a positive prognostic significance in breast cancer [62, 63]. Moreover, these paralog-specific effects are context-dependent, i.e., they may be different in different diseases and in CSC from different diseases. For example, in mesothelioma [64] Notch-2 inhibits the pro-survival effects of Notch-1, while in medulloblastoma [59], Notch-2 stimulates tumorigenesis and Notch-1 inhibits it. Hence, the need to determine in CSC from each disease and possibly disease subtype what the role of individual Notch paralogs is.

2) Ligand diversity

There are several Notch ligands with different functions. Canonical ligands include the Delta and Serrate/Jagged families (Delta-1, -3 and -4 and Jagged-1 and -2 in humans). These have multiple EGF-like repeats, a DSL (Delta-Serrate-Lag2) domain with or without a DOS (Delta-Osm11) domain. Serrate/Jagged family ligands also have a cysteine-rich (CR) motif (Figure 2). It has been suggested that DSL-only ligands lacking DOS domains may function in tandem with DOS-only putative co-ligands such as DLK1 (reviewed in [38]). Non-canonical ligands such as contactins, DNER and MAGP1-2 have been described [38]. While Notch binding to ligands expressed in trans (on contiguous cells) triggers Notch activation, some DSL-containg ligands can also bind Notch in cis (on the same cell), causing cis-inhibition. This is relevant to efforts to target specific ligands using monoclonal antibodies (mAb).

3) Notch glycosylation

The affinity of Notch receptors for specific ligands is controlled by NEC glycosylation. POFUT-1 decorates NEC with fucose residues, to which Fringe enzymes (in humans Lunatic, Radical or Manic Fringe) add N-acetylglucosamine (Figure 2). This regulates the differential affinity of Notch receptors for their ligands. In mammals, Fringe-modified Notch-1 loses its ability to respond to Jagged ligands, while Notch-2 does not [38]. POFUT-1 also has an unrelated chaperone activity that allows folded Notch to be presented at the membrane. Glucosyltransferase RUMI also modifies Notch in Drosophila (reviewed in [38]). Notch glycosylation has not been studied in detail in human cancers or in CSC.

4) ADAM redundancy

The ADAM cleavage step can be catalyzed by at least 2 proteases: ADAM17 (also known as TACE, TNF-α converting enzyme) or ADAM10. Recent evidence suggests that ADAM10 is required for ligand-dependent activation of Notch, while ADAM17 participates in ligand-independent receptor activation [65]. This should be kept in mind when considering the use of ADAM inhibitors to block Notch activation.

5) Multiple γ-secretase isoforms and substrates

γ-secretase, which is targeted by several investigational drugs currently being tested in cancer, is a multisubunit transmembrane aspartyl protease that contains presenilin-1 or -2 (the catalytic subunit) plus APH1, Pen2 and Nicastrin. Since there are 2 presenilin isoforms and at least 2 APH isoforms, there can potentially exist at least four different γ-secretase complexes. There is some evidence, reviewed in [38], that these have different biochemical properties and may contribute differently to Notch activation. In addition to cleaving Notch, γ-secretase catalyzes the regulated intramembranous proteolysis (RIP) of many other proteins [66]. These include ADAM10 [67], putative cancer stem cell marker CD44 [66] and circulating tumor cell (CTC) marker EpCAM [68].

6) Multiple forms of NIC and post-translational modifications regulating its stability

It was originally thought that γ-secretase specifically cleaves Notch before V1744. This led to the production of antibodies recognizing the sequence V1774LLS, at the N-terminus of NIC. However, γ-secretase can cleave Notch at several positions in the 1743-1748 region, with a preference for the L1746-S1747 bond [69]. This means that there are potentially several species of NIC, not all of which are detectable by currently available antibodies. These have different half-lives, with the V1744 being the most stable [38]. However, the intracellular stability and transcriptional activity of NIC is regulated by many factors, including hypoxia [70, 71], prolyl isomerase Pin1 [72] and Nemo-like kinase (NLK) [73]. Thus, a potential role of alternative NICs in cancer cannot be ruled out. Cleavage site selection can be affected by the cellular compartment in which cleavage occurs [38]. This is relevant to the choice of methods to detect NIC in clinical specimens. Undetectable staining with a V1774LLS antibody does not automatically imply lack of Notch activity.

7) Cellular trafficking of Notch receptors and ligands

Ubiquitination, endocytosis and endosome sorting regulate the amount of Notch receptors at the membrane and the balance between ligand-mediated receptor activation, ligand-independent receptor activation and degradation [37, 38]. DSL-containing Notch ligands are ubiquitinated by Mindbomb or Neuralized E3 ligases, endocytosed, “activated” by a poorly understood process and recycled to the membrane in “active” form (Figure 3A). Upon binding, ligand-NEC complexes are trans-endocytosed into ligand-expressing cells, a dynamin-dependent process that separates NEC from NTM [74]. Mono-ubiquitination and endocytosis of Notch as a pre-requisite to activation have been suggested [75], though this remains controversial [37, 38]. The amount of Notch available at the membrane is controlled by several E3 ubiquitin ligases (Itch/AIP4, Cbl, Deltex, Nedd4) that can direct Notch to either lysosomal degradation or recycling (Figure 3C). Deltex forms a complex with Notch and β-arrestin homolog Kurz, leading to Notch ubiquitination and degradation [76]. In the absence of Kurz, Deltex sorts Notch into late endosomes via Rab5, causing Notch activation [77]. In Drosophila, defects in endosome trafficking caused by mutations in ESCRT complex components or in tumor suppressor LGD (Lethal Giant Disc) cause accumulation of Notch in endosomes, where Notch is activated by γ-secretase, causing aberrant cell proliferation. Aquaporin-related anion transporter big-brain (bib), which regulates endosome acidification, is required for Notch signaling [78], perhaps by facilitating the release of cleaved NIC from endosomes [38]. In asymmetrically dividing cells, endocytic mediator Numb, in cooperation with ACBD3, adaptin and Numb-associated kinase (NAK) promotes selective degradation of Notch in one of two daughter cells. This is thought to promote differentiation of the daughter cell that loses Notch. Loss of Numb has been described in breast cancer [79] and asymmetric cell division is a hallmark of “stemness”. Loss of Numb could conceivably result in aberrant retention of “stemness”. Endocytic mediators dynamin and Rab5 are also required in Notch signal-receiving cells. Available evidence indicates that the γ-secretase cleavage step can occur at multiple cellular locations, including at or near the plasma membrane and after endocytosis (reviewed in [38]).

8) Chromatin cross-talk

Finally, though there are many putative CSL responsive elements in the genome, the actual pattern of usage (and thus the gene expression changes affected by Notch activation) appear to depend on the functional interaction of Notch with other nuclear effectors, including for example HIF-1α [70], and the estrogen receptor ERα [52]. Two NTCs can cooperatively bind to sequence-paired CSL-responsive elements [80], possibly making some promoters differentially responsive to monomeric versus dimeric NTC.

These basic observations have significant clinical implications. Simply determining levels of Notch expression at the protein or even mRNA level is not necessarily indicative of how active the pathway is in a specific cancer cell. The best target genes to use as biomarkers may well vary in different diseases and disease subsets. Investigators developing γ-secretase inhibitors (GSIs) should be mindful of possible selectivity for γ-secretase isoforms, of multiple off-target effects and of the fact that compounds with different chemical properties may affect Notch cleavage at the membrane and/or in different intracellular compartments, at neutral or acidic pH. This may directly affect the pharmacological activity of GSIs in vivo.

Targeting Notch in cancer and CSC

Notch signaling has been implicated in a growing number of hematopoietic and solid tumors [39-49]. In most cases, one or more Notch paralogs have oncogenic activity. Inappropriate Notch activation stimulates proliferation, restricts differentiation and/or prevents apoptosis. In the skin, Notch-1 appears to act as a tumor suppressor. Recent evidence suggests that this is an indirect effect. Loss of Notch signaling in the skin causes a barrier defect which causes local inflammation, predisposing to transformation, hyperproduction of thymic stromal lymphopoietin (TSLP), and systemic immunological disturbances [81]. For a general discussion of Notch targeting in cancer, the reader is referred to [35]. This discussion will focus on Notch as a target in CSC.

The strongest evidence to date for a role of Notch in CSC is in breast cancer [6, 8, 82-84], embryonal brain tumors [85], and gliomas [86, 87]. GSIs abolish the formation of secondary mammospheres from a variety of human breast cancer cell lines as well as primary patient specimens (Grudzien et al., submitted for publication, Figure 4). In breast ductal carcinoma in situ (DCIS), the ability to form multilineage spheroids (“mammospheres”, an indicator of stem-like cells) is dramatically decreased by GSIs, a Notch-4 monoclonal antibody or gefitinib [83]. This suggests cooperation between EGFR and Notch-4 in DCIS “stem cell” maintenance. There is evidence for a feedback loop between Her2/Neu and Notch [88, 89], which may maintain CSC in Her2/Neu-overexpressing tumors [90]. Sansone et al. [91] showed that in mammospheres from human breast cancers, interleukin-6 (IL-6) induces Notch-3 signaling, increases expression of Jagged-1 and, through Notch-3, promotes a hypoxia-resistant phenotype. The same group [84] described a p66Shc-Notch-3 pathway as essential to maintain the hypoxia-resistant phenotype of human breast cancer mammospheres. Fan et al. [85] showed that Notch inhibition selectively depletes medulloblastoma CSC as determined by CD133-high status or dye exclusion. The same group has described very similar findings in glioblastoma CSC [86]. Importantly, in gliomas Notch appears to confer radio-resistance to CSC [87]. GSI treatment selectively enhanced radiation-induced death of glioma CSC but not bulk glioma cells. This effect was replicated by Notch-1 or Notch-2 knockdown, and was accompanied by AKT inhibition and reduced and Mcl-1 expression. Other malignancies are being actively investigated. A role of Notch, STAT3 and TGF-β in hepatocellular carcinoma CSC maintenance has been suggested [92]. In gemcitabine-resistant pancreatic carcinoma cells, EMT is associated with activation of Notch signaling, potentially linking Notch to the “Weinberg model” of stemness acquisition through EMT [93] and to treatment resistance. Inhibition of Notch signaling through GSIs [86] or Delta-4 mAb [94] decreased the numbers of CSC and/or their tumorigenicity in some preclinical models.

Figure 4
Effects of a GSI on human breast cancer secondary mammospheres

These encouraging results suggest that therapeutic regimens including Notch inhibitors may be used in the clinic to target CSC and reverse or prevent chemo- or radio-resistance. However, for clinicians interested in targeting Notch in CSC, numerous questions remain to be addressed. There are numerous investigational Notch inhibitors to choose from, some of which are already in the clinic (Table 1). At least four chemically distinct GSIs are being developed by pharmaceutical companies. It is unclear whether these drugs are pharmacologically equivalent and, based on our evolving understanding of γ-secretase, it is possible that they may have significant differences in both Notch inhibition and off-target effects. Due to the very nature of their target, GSIs should block the cleavage of all four Notch paralogs, and multiple other γ-secretase substrates. Thus, they are relatively non-selective drugs, although in vivo their dose-limiting toxicity (secretory diarrhea) is due to Notch inhibition in intestinal stem cells. Inhibition of other γ-secretase targets may either contribute to therapeutic efficacy or hinder it, and these off-target effects may be different for different GSIs. Recently, Watters et al. [95] generated a library of murine mammary tumor models from genetically engineered oncogenic mammary stem cells, essentially artificial CSC. These authors treated tumors with a specific GSI (from Merck), and describe a “GSI response signature”, which is enriched for Notch pathway genes but also includes genes from G-protein activated pathways, eicosanoid pathways and others. Which of these effects are secondary to Notch inhibition and which are off-target remains to be established. However, these data provide a useful platform to compare different GSIs, and GSIs to other agents. The discovery that some NSAIDs-related compounds can allosterically modify the substrate specificity of γ-secretase [96] suggests that Notch-selective γ-secretase modifiers (GSM) can be developed, though specificity for individual Notch homologs remains unlikely. Inhibitors of the MAML/CSL/Notch complex formation were a theoretical possibility until recently. Innovative work by Moellering et al. [97] shows that a hydrocarbon-stapled cell-permeable peptide derived from MAML-1 can selectively prevent the assembly of the NTC and has efficacy as a Notch inhibitor in vitro and in vivo. If non-peptide drugs with similar characteristics can be developed, they would likely be pan-Notch inhibitors without the off-target effects of GSIs. Conversely, mAbs to Notch ligands and receptors offer single-target specificity. Antibodies that “lock” Notch receptors in an inactive conformation by preventing NEC-NTM dissociation are in preclinical development [98]. To date, Notch-1 mAb appear to have similar toxicities compared to GSIs. Delta-4 mAbs are effective against breast CSC in vivo [94]. However, their chronic use can cause vascular neoplasms [99].

Table 1
Notch-targeting agents

GSIs and other small molecules have the advantages of relative ease of administration, oral bioavailability and low cost. Pan-Notch inhibition may or may not be an advantage depending on the relative roles of specific Notch paralogs in individual cancers, and on whether redundancy between Notch paralogs can result in resistance to single-target agents [35]. Off-target effects are not necessarily a disadvantage if they contribute to efficacy. On the other hand, clinical experience so far has shown that GSI must be administered in intermittent dosing regimens to prevent dose-limiting intestinal toxicity. Whether intermittent Notch inhibition is sufficient for effective CSC targeting in patients is an open -and very important- question. If continuous inhibition is necessary for optimal results, systemically delivered GSIs may be at a disadvantage over more specific agents such as anti-ligand mAbs, or may have to be delivered selectively to tumors using innovative pharmaceutics. Situations in which a specific Notch-ligand pair is involved in CSC self-renewal may be targeted more specifically using a mAb. Tumor-selective mAbs (e.g., bispecific antibodies) would theoretically offer site-specific inhibition that could be used chronically. However, the choice of mAb for each specific indication will depend upon the Notch receptors and ligands that play a predominant role in that malignancy's CSC.

Rational combinations and personalized medicine

Even targeting developmental pathways such as Notch will most likely not give us the elusive “magic bullet”, and will require the development of rational combinations. Such combinations will be made possible only through a thorough understanding of cross-talk between Notch and other developmental and non-developmental pathways that may play roles in CSC in specific malignancies. Our knowledge is rapidly evolving, but there is evidence to support some combinations. The following examples are not meant to be all-inclusive, but these classes of agents are reasonable candidates for combination with Notch inhibitors: 1) Inhibitors of the PI3-kinase-AKT-mTOR pathway [35, 87, 100]; 2) NF-κB inhibitors [50, 51, 101], 3) Her2/Neu inhibitors [88, 90], platinum compounds [51, 102], EGFR inhibitors [83] and Hedgehog inhibitors [103]. In breast cancer, a newly discovered feedback between Notch and ERα [52, 104] supports combining Notch inhibitors with anti-estrogens. Anti-estrogen plus GSI and Hedgehog-inhibitor plus GSI combinations are being investigated in ongoing clinical trials. In the case of the Hedgehog inhibitor-GSI combination, anti-CSC effects are being measured specifically.

Ultimately, the best use of Notch inhibitors and other CSC-targeted agents will be in the context of personalized medicine. To that end, we will have to determine: 1) which cancers and specific cancer subtypes contain Notch-dependent CSC; 2) what role do specific components of Notch signaling play in these CSC; 3) what pathways cross-talk with Notch in specific CSCs; and 4) how can one measure Notch activity in CSC from individual patients (e.g., in biopsy material).

The design of clinical trials of CSC-targeted agents will have to consider that anti-CSC effects will not necessarily translate into rapid tumor volume changes. Disease-free or recurrence-free survival will be the most informative endpoints. For situations when this would require prohibitively long follow-ups, it will be important to develop accurate surrogate biomarkers that reflect anti-CSC effects. These may be spheroid formation assays, flow cytometry, molecular tests, or other tests, but post-treatment tumor tissue will be required in most cases. A question of potentially great interest is whether it is possible to assess CSC numbers or the relative “stemness” of individual tumors by studying circulating tumor cells (CTC) [105-107]. These can be isolated from patient blood by several methods, one of which is FDA-approved. While these trials may be challenging, the payoff may be novel treatments that eliminate or greatly reduce treatment-resistance in a whole range of malignancies.


This work was supported by NIH grant P01 AG2553101 and DOD IDEA grant W81XWH-04-1-0478.

Reference List

1. Al Hajj M, Becker MW, Wicha M, Weissman I, Clarke MF. Therapeutic implications of cancer stem cells. Curr Opin Genet Dev. 2004;14:43–7. [PubMed]
2. Alison MR, Islam S, Lim SM. Number crunching in the cancer stem cell market. Breast Cancer Res. 2009;11:302. [PMC free article] [PubMed]
3. Donnenberg VS, Donnenberg AD. Multiple drug resistance in cancer revisited: the cancer stem cell hypothesis. J Clin Pharmacol. 2005;45:872–7. [PubMed]
4. Haraguchi N, Inoue H, Tanaka F, et al. Cancer stem cells in human gastrointestinal cancers. Hum Cell. 2006;19:24–9. [PubMed]
5. Hemmati HD, Nakano I, Lazareff JA, et al. Cancerous stem cells can arise from pediatric brain tumors. Proc Natl Acad Sci U S A. 2003;100:15178–83. [PMC free article] [PubMed]
6. Kakarala M, Wicha MS. Cancer stem cells: implications for cancer treatment and prevention. Cancer J. 2007;13:271–5. [PubMed]
7. Kondo T. Stem cell-like cancer cells in cancer cell lines. Cancer Biomark. 2007;3:245–50. [PubMed]
8. Korkaya H, Wicha MS. Selective targeting of cancer stem cells: a new concept in cancer therapeutics. BioDrugs. 2007;21:299–310. [PubMed]
9. Setoguchi T, Taga T, Kondo T. Cancer stem cells persist in many cancer cell lines. Cell Cycle. 2004;3:414–5. [PubMed]
10. Singh SK, Clarke ID, Terasaki M, et al. Identification of a cancer stem cell in human brain tumors. Cancer Res. 2003;63:5821–8. [PubMed]
11. Song LL, Miele L. Cancer stem cells--an old idea that's new again: implications for the diagnosis and treatment of breast cancer. Expert Opin Biol Ther. 2007;7:431–8. [PubMed]
12. Wang J, Guo LP, Chen LZ, Zeng YX, Lu SH. Identification of cancer stem cell-like side population cells in human nasopharyngeal carcinoma cell line. Cancer Res. 2007;67:3716–24. [PubMed]
13. Yang ZJ, Wechsler-Reya RJ. Hit 'em where they live: targeting the cancer stem cell niche. Cancer Cell. 2007;11:3–5. [PubMed]
14. Foreman KE, Rizzo P, Osipo C, Miele L. The Cancer Stem Cell Hypothesis. In: Bagley R, Teicher B, editors. Stem Cells and Cancer (Cancer Drug Discovery) Springer; New York: 2009. pp. 3–14.
15. Singh SK, Hawkins C, Clarke ID, et al. Identification of human brain tumour initiating cells. Nature. 2004;432:396–401. [PubMed]
16. Eramo A, Lotti F, Sette G, et al. Identification and expansion of the tumorigenic lung cancer stem cell population. Cell Death Differ. 2008;15:504–14. [PubMed]
17. Ricci-Vitiani L, Lombardi DG, Pilozzi E, et al. Identification and expansion of human colon-cancer-initiating cells. Nature. 2007;445:111–5. [PubMed]
18. Collins AT, Berry PA, Hyde C, Stower MJ, Maitland NJ. Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res. 2005;65:10946–51. [PubMed]
19. Yu J, Vodyanik MA, Smuga-Otto K, et al. Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007;318:1917–20. [PubMed]
20. Takahashi K, Tanabe K, Ohnuki M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131:861–72. [PubMed]
21. Sheng XR, Matunis EL. Make room for dedifferentiation. Fly (Austin ) 2009;3:283–5. [PMC free article] [PubMed]
22. Brawley C, Matunis E. Regeneration of male germline stem cells by spermatogonial dedifferentiation in vivo. Science. 2004;304:1331–4. [PubMed]
23. Sheng XR, Brawley CM, Matunis EL. Dedifferentiating spermatogonia outcompete somatic stem cells for niche occupancy in the Drosophila testis. Cell Stem Cell. 2009;5:191–203. [PMC free article] [PubMed]
24. Hugo H, Ackland ML, Blick T, et al. Epithelial--mesenchymal and mesenchymal--epithelial transitions in carcinoma progression. J Cell Physiol. 2007;213:374–83. [PubMed]
25. Mani SA, Guo W, Liao MJ, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 2008;133:704–15. [PMC free article] [PubMed]
26. Dontu G, El Ashry D, Wicha MS. Breast cancer, stem/progenitor cells and the estrogen receptor. Trends Endocrinol Metab. 2004;15:193–7. [PubMed]
27. O'Brien CA, Kreso A, Jamieson C. Cancer stem cells and self-renewal. Clin Cancer Res. 2010;16 in press. [PubMed]
28. Schatton T, Murphy GF, Frank NY, et al. Identification of cells initiating human melanomas. Nature. 2008;451:345–9. [PMC free article] [PubMed]
29. Quintana E, Shackleton M, Sabel MS, Fullen DR, Johnson TM, Morrison SJ. Efficient tumour formation by single human melanoma cells. Nature. 2008;456:593–8. [PMC free article] [PubMed]
30. LaBarge MA. The difficulty of therapeutically targeting cancer stem cell niches. Clin Cancer Res. 2010;16 in press. [PMC free article] [PubMed]
31. Li X, Lewis MT, Huang J, et al. Intrinsic resistance of tumorigenic breast cancer cells to chemotherapy. J Natl Cancer Inst. 2008;100:672–9. [PubMed]
32. Creighton CJ, Li X, Landis M, et al. Residual breast cancers after conventional therapy display mesenchymal as well as tumor-initiating features. Proc Natl Acad Sci U S A. 2009;106:13820–5. [PMC free article] [PubMed]
33. Takahashi-Yanaga F, Kahn M. Targeting Wnt Signaling; Can We Safely Eradicate Cancer Stem Cells?. Clin Cancer Res. 2010;16 in press. [PubMed]
34. Merchant A, Matsui W. Targeting Hedgehog - a Cancer Stem Cell Pathway. Clin Cancer Res. 2010;16 in press. [PMC free article] [PubMed]
35. Rizzo P, Osipo C, Foreman KE, Golde TE, Osborne BA, Miele L. Rational targeting of Notch signaling in cancer. Oncogene. 2008;27:5124–31. [PubMed]
36. Fortini ME, Bilder D. Endocytic regulation of Notch signaling. Curr Opin Genet Dev. 2009;19:323–8. [PMC free article] [PubMed]
37. Fortini ME. Notch signaling: the core pathway and its posttranslational regulation. Dev Cell. 2009;16:633–47. [PubMed]
38. Kopan R, Ilagan MX. The canonical Notch signaling pathway: unfolding the activation mechanism. Cell. 2009;137:216–33. [PMC free article] [PubMed]
39. Allenspach EJ, Maillard I, Aster JC, Pear WS. Notch signaling in cancer. Cancer Biol Ther. 2002;1:466–76. [PubMed]
40. Koch U, Radtke F. Notch and cancer: a double-edged sword. Cell Mol Life Sci. 2007;64:2746–62. [PubMed]
41. Berman JN, Look AT. Targeting transcription factors in acute leukemia in children. Curr Drug Targets. 2007;8:727–37. [PubMed]
42. Shih I, Wang TL. Notch signaling, gamma-secretase inhibitors, and cancer therapy. Cancer Res. 2007;67:1879–82. [PubMed]
43. Roy M, Pear WS, Aster JC. The multifaceted role of Notch in cancer. Curr Opin Genet Dev. 2007;17:52–9. [PubMed]
44. Miele L. Notch signaling. Clin Cancer Res. 2006;12:1074–9. [PubMed]
45. Miele L, Golde T, Osborne B. Notch signaling in cancer. Curr Mol Med. 2006;6:905–18. [PubMed]
46. Purow B. Notch inhibitors as a new tool in the war on cancer: a pathway to watch. Curr Pharm Biotechnol. 2009;10:154–60. [PMC free article] [PubMed]
47. Wang Z, Li Y, Banerjee S, Sarkar FH. Exploitation of the Notch signaling pathway as a novel target for cancer therapy. Anticancer Res. 2008;28:3621–30. [PubMed]
48. Leong KG, Gao WQ. The Notch pathway in prostate development and cancer. Differentiation. 2008;76:699–716. [PubMed]
49. Bailey JM, Singh PK, Hollingsworth MA. Cancer metastasis facilitated by developmental pathways: Sonic hedgehog, Notch, and bone morphogenic proteins. J Cell Biochem. 2007;102:829–39. [PubMed]
50. Vilimas T, Mascarenhas J, Palomero T, et al. Targeting the NF-kappaB signaling pathway in Notch1-induced T-cell leukemia. Nat Med. 2007;13:70–7. [PubMed]
51. Song LL, Peng Y, Yun J, et al. Notch-1 associates with IKKalpha and regulates IKK activity in cervical cancer cells. Oncogene. 2008;27:5833–44. [PubMed]
52. Hao L, Rizzo P, Osipo C, et al. Notch-1 activates estrogen receptor-alpha-dependent transcription via IKKalpha in breast cancer cells. Oncogene. 2010;29:201–13. [PubMed]
53. Guan E, Wang J, Laborda J, Norcross M, Baeuerle PA, Hoffman T. T cell leukemia-associated human Notch/translocation-associated Notch homologue has I kappa B-like activity and physically interacts with nuclear factor-kappa B proteins in T cells. J Exp Med. 1996;183:2025–32. [PMC free article] [PubMed]
54. Wang J, Shelly L, Miele L, Boykins R, Norcross MA, Guan E. Human Notch-1 inhibits NF-kappa B activity in the nucleus through a direct interaction involving a novel domain. J Immunol. 2001;167:289–95. [PubMed]
55. Vacca A, Felli MP, Palermo R, et al. Notch3 and pre-TCR interaction unveils distinct NF-kappaB pathways in T-cell development and leukemia. EMBO J. 2006;25:1000–8. [PMC free article] [PubMed]
56. Sade H, Krishna S, Sarin A. The anti-apoptotic effect of Notch-1 requires p56lck-dependent, Akt/PKB-mediated signaling in T cells. J Biol Chem. 2004;279:2937–44. [PubMed]
57. Perumalsamy LR, Nagala M, Sarin A. Notch-activated signaling cascade interacts with mitochondrial remodeling proteins to regulate cell survival. Proc Natl Acad Sci U S A. 2010 [PMC free article] [PubMed]
58. Bigas A, Martin DI, Milner LA. Notch1 and Notch2 inhibit myeloid differentiation in response to different cytokines. Mol Cell Biol. 1998;18:2324–33. [PMC free article] [PubMed]
59. Fan X, Mikolaenko I, Elhassan I, et al. Notch1 and notch2 have opposite effects on embryonal brain tumor growth. Cancer Res. 2004;64:7787–93. [PubMed]
60. Shimizu K, Chiba S, Saito T, Kumano K, Hamada Y, Hirai H. Functional diversity among Notch1, Notch2, and Notch3 receptors. Biochem Biophys Res Commun. 2002;291:775–9. [PubMed]
61. Nefedova Y, Cheng P, Alsina M, Dalton WS, Gabrilovich DI. Involvement of Notch-1 signaling in bone marrow stroma-mediated de novo drug resistance of myeloma and other malignant lymphoid cell lines. Blood. 2004;103:3503–10. [PubMed]
62. O'Neill CF, Urs S, Cinelli C, et al. Notch2 signaling induces apoptosis and inhibits human MDA-MB-231 xenograft growth. Am J Pathol. 2007;171:1023–36. [PMC free article] [PubMed]
63. Parr C, Watkins G, Jiang WG. The possible correlation of Notch-1 and Notch-2 with clinical outcome and tumour clinicopathological parameters in human breast cancer. Int J Mol Med. 2004;14:779–86. [PubMed]
64. Graziani I, Eliasz S, De Marco MA, et al. Opposite effects of Notch-1 and Notch-2 on mesothelioma cell survival under hypoxia are exerted through the Akt pathway. Cancer Res. 2008;68:9678–85. [PubMed]
65. Bozkulak EC, Weinmaster G. Selective use of ADAM10 and ADAM17 in activation of Notch1 signaling. Mol Cell Biol. 2009;29:5679–95. [PMC free article] [PubMed]
66. Kopan R, Ilagan MX. Gamma-secretase: proteasome of the membrane? Nat Rev Mol Cell Biol. 2004;5:499–504. [PubMed]
67. Tousseyn T, Thathiah A, Jorissen E, et al. ADAM10, the rate-limiting protease of regulated intramembrane proteolysis of Notch and other proteins, is processed by ADAMS-9, ADAMS-15, and the gamma-secretase. J Biol Chem. 2009;284:11738–47. [PMC free article] [PubMed]
68. Maetzel D, Denzel S, Mack B, et al. Nuclear signalling by tumour-associated antigen EpCAM. Nat Cell Biol. 2009;11:162–71. [PubMed]
69. Tagami S, Okochi M, Yanagida K, et al. Regulation of Notch signaling by dynamic changes in the precision of S3 cleavage of Notch-1. Mol Cell Biol. 2008;28:165–76. [PMC free article] [PubMed]
70. Gustafsson MV, Zheng X, Pereira T, et al. Hypoxia requires notch signaling to maintain the undifferentiated cell state. Dev Cell. 2005;9:617–28. [PubMed]
71. Chen Y, De Marco MA, Graziani I, et al. Oxygen concentration determines the biological effects of NOTCH-1 signaling in adenocarcinoma of the lung. Cancer Res. 2007;67:7954–9. [PubMed]
72. Rustighi A, Tiberi L, Soldano A, et al. The prolyl-isomerase Pin1 is a Notch1 target that enhances Notch1 activation in cancer. Nat Cell Biol. 2009;11:133–42. [PubMed]
73. Ishitani T, Hirao T, Suzuki M, et al. Nemo-like kinase suppresses Notch signalling by interfering with formation of the Notch active transcriptional complex. Nat Cell Biol. 2010;12:278–85. [PubMed]
74. Parks AL, Klueg KM, Stout JR, Muskavitch MA. Ligand endocytosis drives receptor dissociation and activation in the Notch pathway. Development. 2000;127:1373–85. [PubMed]
75. Gupta-Rossi N, Six E, LeBail O, et al. Monoubiquitination and endocytosis direct gamma-secretase cleavage of activated Notch receptor. J Cell Biol. 2004;166:73–83. [PMC free article] [PubMed]
76. Mukherjee A, Veraksa A, Bauer A, Rosse C, Camonis J, Artavanis-Tsakonas S. Regulation of Notch signalling by non-visual beta-arrestin. Nat Cell Biol. 2005;7:1191–201. [PubMed]
77. Hori K, Fostier M, Ito M, et al. Drosophila deltex mediates suppressor of Hairless-independent and late-endosomal activation of Notch signaling. Development. 2004;131:5527–37. [PubMed]
78. Kanwar R, Fortini ME. The big brain aquaporin is required for endosome maturation and notch receptor trafficking. Cell. 2008;133:852–63. [PMC free article] [PubMed]
79. Pece S, Serresi M, Santolini E, et al. Loss of negative regulation by Numb over Notch is relevant to human breast carcinogenesis. J Cell Biol. 2004;167:215–21. [PMC free article] [PubMed]
80. Nam Y, Sliz P, Pear WS, Aster JC, Blacklow SC. Cooperative assembly of higher-order Notch complexes functions as a switch to induce transcription. Proc Natl Acad Sci U S A. 2007;104:2103–8. [PMC free article] [PubMed]
81. Demehri S, Morimoto M, Holtzman MJ, Kopan R. Skin-derived TSLP triggers progression from epidermal-barrier defects to asthma. PLoS Biol. 2009;7:e1000067. [PMC free article] [PubMed]
82. Farnie G, Clarke RB. Mammary stem cells and breast cancer--role of Notch signalling. Stem Cell Rev. 2007;3:169–75. [PubMed]
83. Farnie G, Clarke RB, Spence K, et al. Novel cell culture technique for primary ductal carcinoma in situ: role of Notch and epidermal growth factor receptor signaling pathways. J Natl Cancer Inst. 2007;99:616–27. [PubMed]
84. Sansone P, Storci G, Giovannini C, et al. p66Shc/Notch-3 interplay controls self-renewal and hypoxia survival in human stem/progenitor cells of the mammary gland expanded in vitro as mammospheres. Stem Cells. 2007;25:807–15. [PubMed]
85. Fan X, Matsui W, Khaki L, et al. Notch Pathway Inhibition Depletes Stem-like Cells and Blocks Engraftment in Embryonal Brain Tumors. Cancer Res. 2006;66:7445–52. [PubMed]
86. Fan X, Khaki L, Zhu TS, et al. Notch Pathway Blockade Depletes CD133-Positive Glioblastoma Cells and Inhibits Growth of Tumor Neurospheres and Xenografts. Stem Cells. 2010;28:5–16. [PMC free article] [PubMed]
87. Wang J, Wakeman TP, Lathia JD, et al. Notch Promotes Radioresistance of Glioma Stem Cells. Stem Cells. 2010;28:17–28. [PMC free article] [PubMed]
88. Osipo C, Patel P, Rizzo P, et al. ErbB-2 inhibition activates Notch-1 and sensitizes breast cancer cells to a gamma-secretase inhibitor. Oncogene. 2008;27:5019–32. [PubMed]
89. Chen Y, Fischer WH, Gill GN. Regulation of the ERBB-2 promoter by RBPJkappa and NOTCH. J Biol Chem. 1997;272:14110–4. [PubMed]
90. Korkaya H, Wicha MS. HER-2, notch, and breast cancer stem cells: targeting an axis of evil. Clin Cancer Res. 2009;15:1845–7. [PubMed]
91. Sansone P, Storci G, Tavolari S, et al. IL-6 triggers malignant features in mammospheres from human ductal breast carcinoma and normal mammary gland. J Clin Invest. 2007;117:3988–4002. [PMC free article] [PubMed]
92. Yao Z, Mishra L. Cancer stem cells and hepatocellular carcinoma. Cancer Biol Ther. 2009;8:1691–8. [PMC free article] [PubMed]
93. Wang Z, Li Y, Kong D, et al. Acquisition of epithelial-mesenchymal transition phenotype of gemcitabine-resistant pancreatic cancer cells is linked with activation of the notch signaling pathway. Cancer Res. 2009;69:2400–7. [PMC free article] [PubMed]
94. Hoey T, Yen WC, Axelrod F, et al. DLL4 blockade inhibits tumor growth and reduces tumor-initiating cell frequency. Cell Stem Cell. 2009;5:168–77. [PubMed]
95. Watters JW, Cheng C, Majumder PK, et al. De novo discovery of a gamma-secretase inhibitor response signature using a novel in vivo breast tumor model. Cancer Res. 2009;69:8949–57. [PubMed]
96. Kukar T, Golde TE. Possible mechanisms of action of NSAIDs and related compounds that modulate gamma-secretase cleavage. Curr Top Med Chem. 2008;8:47–53. [PMC free article] [PubMed]
97. Moellering RE, Cornejo M, Davis TN, et al. Direct inhibition of the NOTCH transcription factor complex. Nature. 2009;462:182–8. [PMC free article] [PubMed]
98. Li K, Li Y, Wu W, et al. Modulation of notch signaling by antibodies specific for the extracellular negative regulatory region of Notch3. J Biol Chem. 2008;283:8046–54. [PubMed]
99. Yan M, Callahan CA, Beyer JC, et al. Chronic DLL4 blockade induces vascular neoplasms. Nature. 2010;463:E6–E7. [PubMed]
100. Meurette O, Stylianou S, Rock R, Collu GM, Gilmore AP, Brennan K. Notch activation induces Akt signaling via an autocrine loop to prevent apoptosis in breast epithelial cells. Cancer Res. 2009;69:5015–22. [PubMed]
101. Osipo C, Golde TE, Osborne BA, Miele LA. Off the beaten pathway: the complex cross talk between Notch and NF-kappaB. Lab Invest. 2008;88:11–7. [PubMed]
102. Meng RD, Shelton CC, Li YM, et al. gamma-Secretase inhibitors abrogate oxaliplatin-induced activation of the Notch-1 signaling pathway in colon cancer cells resulting in enhanced chemosensitivity. Cancer Res. 2009;69:573–82. [PMC free article] [PubMed]
103. Wang Z, Li Y, Banerjee S, Sarkar FH. Emerging role of Notch in stem cells and cancer. Cancer Lett. 2009;279:8–12. [PMC free article] [PubMed]
104. Rizzo P, Miao H, D'Souza G, et al. Cross-talk between notch and the estrogen receptor in breast cancer suggests novel therapeutic approaches. Cancer Res. 2008;68:5226–35. [PubMed]
105. Budd GT. Let me do more than count the ways: what circulating tumor cells can tell us about the biology of cancer. Mol Pharm. 2009;6:1307–10. [PubMed]
106. Jacob K, Sollier C, Jabado N. Circulating tumor cells: detection, molecular profiling and future prospects. Expert Rev Proteomics. 2007;4:741–56. [PubMed]
107. Paterlini-Brechot P, Benali NL. Circulating tumor cells (CTC) detection: clinical impact and future directions. Cancer Lett. 2007;253:180–204. [PubMed]
108. Cordle J, Johnson S, Tay JZ, et al. A conserved face of the Jagged/Serrate DSL domain is involved in Notch trans-activation and cis-inhibition. Nat Struct Mol Biol. 2008;15:849–57. [PMC free article] [PubMed]
109. Nam Y, Weng AP, Aster JC, Blacklow SC. Structural requirements for assembly of the CSL/intracellular notch1/mastermind-like 1 transcriptional activation complex. J Biol Chem. 2003;278:21232–9. [PubMed]
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