IGFBP7 induces apoptosis of acute myeloid leukemia cells and synergizes with chemotherapy in suppression of leukemia cell survival

Despite high remission rates after chemotherapy, only 30–40% of acute myeloid leukemia (AML) patients survive 5 years after diagnosis. This extremely poor prognosis of AML is mainly caused by treatment failure due to chemotherapy resistance. Chemotherapy resistance can be caused by various features including activation of alternative signaling pathways, evasion of cell death or activation of receptor tyrosine kinases such as the insulin growth factor-1 receptor (IGF-1R). Here we have studied the role of the insulin-like growth factor-binding protein-7 (IGFBP7), a tumor suppressor and part of the IGF-1R axis, in AML. We report that IGFBP7 sensitizes AML cells to chemotherapy-induced cell death. Moreover, overexpression of IGFBP7 as well as addition of recombinant human IGFBP7 is able to reduce the survival of AML cells by the induction of a G2 cell cycle arrest and apoptosis. This effect is mainly independent from IGF-1R activation, activated Akt and activated Erk. Importantly, AML patients with high IGFBP7 expression have a better outcome than patients with low IGFBP7 expression, indicating a positive role for IGFBP7 in treatment and outcome of AML. Together, this suggests that the combination of IGFBP7 and chemotherapy might potentially overcome conventional AML drug resistance and thus might improve AML patient survival.Only 30–40% of acute myeloid leukemia (AML) patients survive 5 years after diagnosis.¹ This extremely poor prognosis is mainly caused by treatment failure due to chemotherapy resistance. This resistance is often a multifactorial phenomenon that can include enhanced expression or activation of receptor tyrosine kinases such as the insulin growth factor-1 receptor (IGF-1R).^{2, 3} The IGF-1R stimulates proliferation, protects cells from apoptosis and has been implicated in the development and maintenance of various cancers.^{4, 5} Several oncogenes require an intact IGF-1R pathway for their transforming activity⁶ and moreover, disruption or inhibition of IGF-1R activity has been shown to inhibit the growth and motility of a wide range of cancer cells in vitro and in mouse models.^{4, 5} IGF-1Rs are membrane receptors and binding of their ligand, the insulin-like growth factor-1 (IGF-1), results in receptor phosphorylation and activation of MAPK and PI3K/Akt signaling.⁴ Importantly, IGF-1, normally produced by the liver and bone marrow stromal cells, can stimulate the proliferation of cancer cells in vitro and genetic manipulations that reduce IGF-1 signaling can lead to decreased tumor growth.^{7, 8}In hematological malignancies, a role for IGF-1 signaling has been demonstrated in multiple myeloma (MM) where it stimulates growth and potently mediates survival.⁹ Several anti-IGF-1R strategies have been shown to inhibit MM growth.^{10, 11} In AML, expression of the IGF-1R and IGF-1 was detected in AML cell lines and primary AML blasts and stimulation with IGF-1 can promote the growth of AML cells.^{12, 13, 14} In addition, neutralizing IGF-1R antibodies and the tyrosine kinase inhibitors (TKIs) NVP-AEW541 and NVP-ADW742, have been shown to inhibit proliferation and to induce apoptosis.^{15, 16}In addition to its mitogenic and anti-apoptotic roles, directly influencing tumor development, IGF-1R appears to be a critical determinant of response to numerous anti-cancer therapies, including TKIs and chemotherapy.^{2, 3, 17, 18, 19, 20, 21, 22} In AML, activated IGF-1R signaling has been linked to cytarabine resistance, a drug included in every AML treatment schedule.¹⁷ Notably, in several cancer cell lines, a small subpopulation of drug-tolerant cancer cells exists that maintains their viability, after treatment with a lethal drug dose, via engagement of the IGF-1R.¹⁸The activity of the IGF-1R is tightly controlled at multiple levels, including their processing, endocytosis, trafficking and availability of its ligands.⁴ Ligand bioavailability is partly controlled by the family of secreted insulin-like growth factor-binding protein (IGFBP1 to IGFBP6), which can bind to IGFs therewith regulating the interaction of these ligands to their receptors. However, as IGFBPs are able to induce IGF-dependent and IGF-independent effects, the results of several studies on their role in cancer cell survival appeared to be controversial and complex.^{23, 24} In addition to IGFBPs, various IGFBP-related proteins have been identified.^{23, 25} One of these is the IGFB-related protein 1, also known as insulin-like growth factor-binding protein-7 (IGFBP7). IGFBP7 has 30% homology to IGFBP1 to IGFBP6 in its N-terminal domain and functions predominantly as a tumor suppressor.^{23, 24, 25, 26} In contrast to IGFBP1 to IGFBP6, which bind to the IGFs,²³ IGFBP7 is a secreted protein that can directly bind to the IGF-1R and thereby inhibits its activity.²⁷ The abundance of IGFBP7 is inversely correlated with tumor progression in hepatocellular carcinoma.²⁸ Importantly, decreased expression of IGFBP7 has been associated with therapy resistance^{29, 30} and increasing IGFBP7 levels can inhibit melanoma and breast cancer growth.^{31, 32} IGFBP7 was originally identified as being involved in Raf-mediated apoptosis and senescence³³ and also has been shown to induce senescence in mesenchymal stromal cells.³⁴We established that IGFBP7 induces a cell cycle block and apoptosis in AML cells and cooperates with chemotherapy in the induction of leukemia cell death. AML patients with low IGFBP7 expression have a worse outcome than patients with high IGFBP7 expression, indicating that AML patients might benefit from a combination therapy consisting of chemotherapy and IGFBP7. Our results define IGFBP7 as a focus to enhance chemotherapy efficacy and improve AML patient survival.     

Shrub encroachment in North American grasslands: shifts in growth form dominance rapidly alters control of ecosystem carbon inputs

Shrub encroachment into grass-dominated biomes is occurring globally due to a variety of anthropogenic activities, but the consequences for carbon (C) inputs, storage and cycling remain unclear. We studied eight North American graminoid-dominated ecosystems invaded by shrubs, from arctic tundra to Atlantic coastal dunes, to quantify patterns and controls of C inputs via aboveground net primary production (ANPP). Across a fourfold range in mean annual precipitation (MAP), a key regulator of ecosystem C input at the continental scale, shrub invasion decreased ANPP in xeric sites, but dramatically increased ANPP (>1000 g m⁻²) at high MAP, where shrub patches maintained extraordinarily high leaf area. Concurrently, the relationship between MAP and ANPP shifted from being nonlinear in grasslands to linear in shrublands. Thus, relatively abrupt (<50 years) shifts in growth form dominance, without changes in resource quantity, can fundamentally alter continental-scale pattern of C inputs and their control by MAP in ways that exceed the direct effects of climate change alone.     

Ovipositor internal microsculpture and other features in doryctine wasps (Insecta, Hymenoptera, Braconidae)

Detailed ovipositor morphology has been investigated in representatives of 81 of the more than 120 genera of the parasitic wasp subfamily Doryctinae. Potentially phylogenetically informative variation was found in the microsculpture of the egg canal wall and the presence and degree of development of the valvillus. In all the genera, a socketed seta arises from below each member of the line of ctenidia (comb-like scales) distal to the valvillus (or valvillar region). These subctenidial setae are modified in two groups of genera. In the Holcobraconini (= Odontobraconini) together with the genera Acanthodoryctes, Binarea, Monarea , and Liobrucon , the setae are extremely flattened producing overlapping leaflet-like structures. In some of these, groups of leaflets are further modified to form fans that protrude into the egg canal. In Schlettereriella and Leptospathius the basal seta is highly branched and rather flattened and may serve a similar function to the valvillus which in these two genera is extremely reduced. In Doryctes and Neodoryctes , there is a distinctive type of bar-like sculpture anterior to the valvillus and a single large crescentic bar posterior to the valvillus. These modifications are suggested as possible synapomorphies (in lieu of an analysis) for their respective groups of genera. Several other characters are also described and illustrated. CO 1998 The Norwegian Academy of Science and Letters     

Development of multiple strain competitive index assays for <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> using pIMC; a new site-specific integrative vector

Background  

The foodborne, gram-positive pathogen, Listeria monocytogenes, is capable of causing lethal infections in compromised individuals. In the post genomic era of L. monocytogenes research, techniques are required to identify and validate genes involved in the pathogenicity and environmental biology of the organism. The aim here was to develop a widely applicable method to tag L. monocytogenes strains, with a particular emphasis on the development of multiple strain competitive index assays.     

Two new genera and species of Braconinae (Insecta, Hymenoptera, Braconidae)from Brunei

Fuvihracon gen.n. (typc-species: Fuvibracon gauldi sp.n.) and Spurhulibrucon (type-species: Sparhulibracon dayi sp.n.), both lrom Brunei, are described and illustrated. The relationships of the two new genera and of the Carnpyloneurus Szépligeti and Chelonogaslra Ashmead generie groups are discussed.     

Morphology of the antenna cleaner in the Hymenoptera with particular reference to non-aculeate families (Insecta)

An electron microscopic survey of antenna cleaner morphology, mainly in non-aculcate Hymenoptera, is presented. Modified, scale- or paddle-shaped setae on the fore basitarsus were found to be widely distributed throughout the order, but were particularly well developed in the Xyeloidea. Megalodontoidea, Blasticotomidae, Siricoidca, Orussoidea, Cephoidea and Chalcidoidea. as well as in the aculeate family Formicidae. A comb of fine setae on the fore basitarsus was present in all Apocrita, with the exception of the Trigonalyoidea, Evaniidae, and some families of Chalcidoidea, but among the symphytan families was present only in the Orussidae. Members of the symphytan family, Anaxyelidae have a distinct line of discrete setae in the same position as the fine comb of the Orussidae and apocritans which we term a protocomb; members of the Cephidae also show an indication of a protocomb, in the form of a line of more widely spaced, erect setae, that could form part of the same transition series. Members of the Trigonalyoidea and of the Evaniidae have no comb of fine setae but do possess one or more rows of highly modified, plate-like structures on the fore basitarsi.     

Molecular evidence of cycad seed predation by immature Aulacoscelidinae (Coleoptera: Orsodacnidae)

Adult beetles in the small subfamily Aulacoscelidinae (superfamily Chrysomeloidea) are known to feed on the foliage and juices of New World cycads (Order Cycadales; family Zamiaceae), but the habits of larvae have long remained a mystery. We provide the first direct evidence that Aulacoscelidinae larvae feed on and develop within the megagametophyte of the Mesoamerican cycad, Dioon merolae (Zamiaceae). Phylogenetic analyses based on partial DNA sequences from 3 genes recover a cycad seed‐feeding larva proposed to belong to Aulacoscelidinae. These observations reveal a more intimate feeding relationship between Aulacoscelidinae and their New World cycad host plants than was previously recognized. Further, adult Aulacoscelidinae have long been noted to resemble Jurassic fossil chrysomeloids in the extinct subfamily Protoscelidinae. The molecular, morphological, ecological and fossil data reported herein are broadly compatible with an early association between Aulacoscelidinae and their gymnosperm hosts.     

Starch Metabolism in Shoot-forming Tobacco Callus

Starch, free sugars and protein contents, and the specific activitiesof enzymes of starch metabolism were determined in tobacco calluscultured under shoot-forming and non-shoot-forming conditions.Shoot-forming cultures contained higher levels of starch, freesugars and protein. Shoot-forming cultures had higher specificactivities for starch-synthesising enzymes throughout culture.On the other hand, higher levels of activity for starch-degradingenzymes in shoot-forming tissues were only observed during organizeddevelopment. The role of phosphorylase in the cultured tissuewas not clear.