Effectiveness of Carboplatin and Paclitaxel as First- and Second-Line Treatment in 61 Patients with Metastatic Melanoma

BACKGROUND: Patients with metastatic melanoma have a very unfavorable prognosis with few therapeutic options. Based on previous promising experiences within a clinical trial involving carboplatin and paclitaxel a series of advanced metastatic melanoma patients were treated with this combination. METHODS: Data of all patients with cutaneous metastatic melanoma treated with carboplatin and paclitaxel (CP) at our institution between October 2005 and December 2007 were retrospectively evaluated. For all patients a once-every-3-weeks dose-intensified regimen was used. Overall and progression free survival were calculated using the method of Kaplan and Meier. Tumour response was evaluated according to RECIST criteria. RESULTS: 61 patients with cutaneous metastatic melanoma were treated with CP. 20 patients (85% M1c) received CP as first-line treatment, 41 patients (90.2% M1c) had received at least one prior systemic therapy for metastatic disease. Main toxicities were myelosuppression, fatigue and peripheral neuropathy. Partial responses were noted in 4.9% of patients, stable disease in 23% of patients. No complete response was observed. Median progression free survival was 10 weeks. Median overall survival was 31 weeks. Response, progression-free and overall survival were equivalent in first- and second-line patients. 60 patients of 61 died after a median follow up of 7 months. Median overall survival differed for patients with controlled disease (PR+SD) (49 weeks) compared to patients with progressive disease (18 weeks). CONCLUSIONS: Among patients with metastatic melanoma a subgroup achieved disease control under CP therapy which may be associated with a survival benefit. This potential advantage has to be weighed against considerable toxicity. Since response rates and survival were not improved in previously untreated patients compared to pretreated patients, CP should thus not be applied as first-line treatment

Inhibition of STAT3 signaling prevents vascular smooth muscle cell proliferation and neointima formation

Dedifferentiation, migration, and proliferation of resident vascular smooth muscle cells (SMCs) are key components of neointima formation after vascular injury. Activation of signal transducer and activator of transcription-3 (STAT3) is suggested to be critically involved in this process, but the complex regulation of STAT3-dependent genes and the functional significance of inhibiting this pathway during the development of vascular proliferative diseases remain elusive. In this study, we demonstrate that STAT3 was activated in neointimal lesions following wire-induced injury in mice. Phosphorylation of STAT3 induced trans-activation of cyclin D1 and survivin in SMCs in vitro and in neointimal cells in vivo, thus promoting proliferation and migration of SMCs as well as reducing apoptotic cell death. WP1066, a highly potent inhibitor of STAT3 signaling, abrogated phosphorylation of STAT3 and dose-dependently inhibited the functional effects of activated STAT3 in stimulated SMCs. The local application of WP1066 via a thermosensitive pluronic F-127 gel around the dilated arteries significantly inhibited proliferation of neointimal cells and decreased the neointimal lesion size at 3 weeks after injury. Even though WP1066 application attenuated the injury-induced up-regulation of the chemokine RANTES at 6 h after injury, there was no significant effect on the accumulation of circulating cells at 1 week after injury. In conclusion, these data identify STAT3 as a key molecule for the proliferative response of SMC and neointima formation. Moreover, inhibition of STAT3 by the potent and specific compound WP1066 might represent a novel and attractive approach for the local treatment of vascular proliferative diseases

CXCR4 Expression in Prostate Cancer Progenitor Cells

Tumor progenitor cells represent a population of drug-resistant cells that can survive conventional chemotherapy and lead to tumor relapse. However, little is known of the role of tumor progenitors in prostate cancer metastasis. The studies reported herein show that the CXCR4/CXCL12 axis, a key regulator of tumor dissemination, plays a role in the maintenance of prostate cancer stem-like cells. The CXCL4/CXCR12 pathway is activated in the CD44+/CD133+ prostate progenitor population and affects differentiation potential, cell adhesion, clonal growth and tumorigenicity. Furthermore, prostate tumor xenograft studies in mice showed that a combination of the CXCR4 receptor antagonist AMD3100, which targets prostate cancer stem-like cells, and the conventional chemotherapeutic drug Taxotere, which targets the bulk tumor, is significantly more effective in eradicating tumors as compared to monotherapy

Chromatin-associated regulation of sorbitol synthesis in flower buds of peach

[EN] Key message PpeS6PDH gene is postulated to mediate sorbitol synthesis in flower buds of peach concomitantly with specific chromatin modifications. Abstract Perennial plants have evolved an adaptive mechanism involving protection of meristems within specialized structures named buds in order to survive low temperatures and water deprivation during winter. A seasonal period of dormancy further improves tolerance of buds to environmental stresses through specific mechanisms poorly known at the molecular level. We have shown that peach PpeS6PDH gene is down-regulated in flower buds after dormancy release, concomitantly with changes in the methylation level at specific lysine residues of histone H3 (H3K27 and H3K4) in the chromatin around the translation start site of the gene. PpeS6PDH encodes a NADPH-dependent sorbitol-6-phosphate dehydrogenase, the key enzyme for biosynthesis of sorbitol. Consistently, sorbitol accumulates in dormant buds showing higher PpeS6PDH expression. Moreover, PpeS6PDH gene expression is affected by cold and water deficit stress. Particularly, its expression is up-regulated by low temperature in buds and leaves, whereas desiccation treatment induces PpeS6PDH in buds and represses the gene in leaves. These data reveal the concurrent participation of chromatin modification mechanisms, transcriptional regulation of PpeS6PDH and sorbitol accumulation in flower buds of peach. In addition to its role as a major translocatable photosynthate in Rosaceae species, sorbitol is a widespread compatible solute and cryoprotectant, which suggests its participation in tolerance to environmental stresses in flower buds of peach.This work was funded by the Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA)-FEDER (RF2013-00043-C02-02) and the Ministry of Science and Innovation of Spain (AGL2010-20595). AL was funded by a fellowship co-financed by the European Social Fund and the Instituto Valenciano de Investigaciones Agrarias (IVIA).Lloret, A.; Martinez Fuentes, A.; Agustí Fonfría, M.; Badenes, ML.; Rios, G. (2017). Chromatin-associated regulation of sorbitol synthesis in flower buds of peach. Plant Molecular Biology. 95(4-5):507-517. https://doi.org/10.1007/s11103-017-0669-6S507517954-5Andersen CL, Jensen JL, Ørntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64:5245–5250. doi: 10.1158/0008-5472.CAN-04-0496Bai S, Saito T, Ito A et al (2016) Small RNA and PARE sequencing in flower bud reveal the involvement of sRNAs in endodormancy release of Japanese pear (Pyrus pyrifolia ‘Kosui’). BMC Genomics 17:230. doi: 10.1186/s12864-016-2514-8Bielenberg DG, Wang Y, Li Z et al (2008) Sequencing and annotation of the evergrowing locus in peach (Prunus persica [L.] Batsch) reveals a cluster of six MADS-box transcription factors as candidate genes for regulation of terminal bud formation. Tree Genet Genomes 4:495–507. doi: 10.1007/s11295-007-0126-9Bieleski RL (1969) Accumulation and translocation of sorbitol in apple phloem. Aust J Biol Sci 22:611–620. doi: 10.1071/BI9690611Bieleski RL (1982) Sugar alcohols. In: Loewus F, Tanner W (eds) Encyclopedia of plant physiology, new series 13A. Springer-Verlag, Berlin, pp 158–192Bortiri E, Oh SH, Gao FY, Potter D (2002) The phylogenetic utility of nucleotide sequences of sorbitol 6-phosphate dehydrogenase in Prunus (Rosaceae). Am J Bot 89:1697–1708. doi: 10.3732/ajb.89.10.1697Chouard P (1960) Vernalization and its relations to dormancy. Annu Rev Plant Physiol 11:191–238. doi: 10.1146/annurev.pp.11.060160.001203Conde D, Le Gac AL, Perales M et al (2017) Chilling-responsive DEMETER-LIKE DNA demethylase mediates in poplar bud break. Plant Cell Environ 40:2236–2249. doi: 10.1111/pce.13019Couvillon GA, Erez A (1985) Influence of prolonged exposure to chilling temperatures on bud break and heat requirement for bloom of several fruit species. J Amer Soc Hort Sci 110:47–50de la Fuente L, Conesa A, Lloret A, Badenes ML, Ríos G (2015) Genome-wide changes in histone H3 lysine 27 trimethylation associated with bud dormancy release in peach. Tree Genet Genomes 11:45. doi: 10.1007/s11295-015-0869-7Deng W, Buzas DM, Ying H et al (2013) Arabidopsis polycomb repressive complex 2 binding sites contain putative GAGA factor binding motifs within coding regions of genes. BMC Genomics 14:593. doi: 10.1186/1471-2164-14-593Escobar-Gutiérrez AJ, Gaudillère JP (1996) Distribution, metabolism and role of sorbitol in higher plants—A review. Agronomie 16:281–298. doi: 10.1051/agro:19960502Escobar-Gutiérrez AJ, Zipperlin B, Carbonne F, Moing A, Gaudillére JP (1998) Photosynthesis, carbon partitioning and metabolite content during drought stress in peach seedlings. Aust J Plant Physiol 25:197–205. doi: 10.1071/PP97121Eshghi S, Tafazoli E, Dokhani S, Rahemi M, Emam Y (2007) Changes in carbohydrate contents in shoot tips, leaves and roots of strawberry (Fragaria x ananassa Duch) during flower-bud differentiation. Sci Hortic 113:255–260. doi: 10.1016/j.scienta.2007.03.014Everard JD, Cantini C, Grumet R, Plummer J, Loescher WH (1997) Molecular cloning of mannose-6-phosphate reductase and its developmental expression in celery. Plant Physiol 113:1427–1435. doi: 10.1104/pp.113.4.1427Fennell A (2014) Genomics and functional genomics of winter low temperature tolerance in temperate fruit crops. Crit Rev Plant Sci 33:125–140. doi: 10.1080/07352689.2014.870410Figueroa CM, Iglesias AA (2010) Aldose-6-phosphate reductase from apple leaves: importance of the quaternary structure for enzyme activity. Biochimie 92:81–88. doi: 10.1016/j.biochi.2009.09.013Gao M, Tao R, Miura K, Dandekar AM, Sugiura A (2001) Transformation of Japanese persimmon (Diospyros kaki Thunb) with apple cDNA encoding NADP-dependent sorbitol-6-phosphate dehydrogenase. Plant Sci 160:837–845. doi: 10.1016/S0168-9452(00)00458-1Grant CR, ap Rees T (1981) Sorbitol metabolism by apple seedlings. Phytochemistry 20:1505–1511. doi: 10.1016/S0031-9422(00)98521-2Hartman MD, Figueroa CM, Arias DG, Iglesias AA (2017) Inhibition of recombinant aldose-6-phosphate reductase from peach leaves by hexose-phosphates, inorganic phosphate and oxidants. Plant Cell Physiol 58:145–155. doi: 10.1093/pcp/pcw180Horvath DP, Anderson JV, Chao WS, Foley ME (2003) Knowing when to grow: signals regulating bud dormancy. Trends Plant Sci 8:534–540. doi: 10.1016/j.tplants.2003.09.013Horvath DP, Sung S, Kim D, Chao W, Anderson J (2010) Characterization, expression and function of DORMANCY ASSOCIATED MADS-BOX genes from leafy spurge. Plant Mol Biol 73:169–179. doi: 10.1007/s11103-009-9596-5Hussain S, Niu Q, Yang F, Hussain N, Teng Y (2015) The possible role of chilling in floral and vegetative bud dormancy release in Pyrus pyrifolia. Biol Plant 59:726–734. doi: 10.1007/s10535-015-0547-5Hyndman D, Baumanb DR, Herediac VV, Penning TM (2003) The aldo-keto reductase superfamily homepage. Chem Biol Interact 143–144:621–631. doi: 10.1016/S0009-2797(02)00193-XIto A, Sakamoto D, Moriguchi T (2012) Carbohydrate metabolism and its possible roles in endodormancy transition in Japanese pear. Sci Hortic 144:187–194. doi: 10.1016/j.scienta.2012.07.009Ito A, Sugiura T, Sakamoto D, Moriguchi T (2013) Effects of dormancy progression and low-temperature response on changes in the sorbitol concentration in xylem sap of Japanese pear during winter season. Tree Physiol 33:398–408. doi: 10.1093/treephys/tpt021Jung S, Bassett C, Bielenberg DG et al (2015) A standard nomenclature for gene designation in the Rosaceae. Tree Genet Genomes 11:108. doi: 10.1007/s11295-015-0931-5Kanayama Y, Mori H, Imaseki H, Yamaki S (1992) Nucleotide sequence of a cDNA encoding NADP-sorbitol-6-phosphate dehydrogenase from apple. Plant Physiol 100:1607–1608Kanayama Y, Watanabe M, Moriguchi R, Deguchi M, Kanahama K, Yamaki S (2006) Effects of low temperature and abscisic acid on the expression of the sorbitol-6-phosphate dehydrogenase gene in apple leaves. J Japan Soc Hort Sci 75:20–25. doi: 10.2503/jjshs.75.20Kumar G, Rattan UK, Singh AK (2016a) Chilling-mediated DNA methylation changes during dormancy and its release reveal the importance of epigenetic regulation during winter dormancy in apple (Malus x domestica Borkh). PLoS ONE 11:e0149934. doi: 10.1371/journal.pone.0149934Kumar S, Stecher G, Tamura K (2016b) MEGA7: molecular evolutionary genetics analysis version 70 for bigger datasets. Mol Biol Evol 33:1870–1874. doi: 10.1093/molbev/msw054Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685. doi: 10.1038/227680a0Larkin MA, Blackshields G, Brown NP et al (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948. doi: 10.1093/bioinformatics/btm404Leida C, Terol J, Martí G et al (2010) Identification of genes associated with bud dormancy release in Prunus persica by suppression subtractive hybridization. Tree Physiol 30:655–666. doi: 10.1093/treephys/tpq008Leida C, Conesa A, Llácer G, Badenes ML, Ríos G (2012) Histone modifications and expression of DAM6 gene in peach are modulated during bud dormancy release in a cultivar-dependent manner. New Phytol 193:67–80. doi: 10.1111/j.1469-8137.2011.03863.xLiang D, Cui M, Wu S, Ma F-W (2012) Genomic structure, sub-cellular localization, and promoter analysis of the gene encoding sorbitol-6-phosphate dehydrogenase from apple. Plant Mol Biol Rep 30:904–914. doi: 10.1007/s11105-011-0409-zLiu D, Ni J, Wu R, Teng Y (2013) High temperature alters sorbitol metabolism in Pyrus pyrifolia leaves and fruit flesh during late stages of fruit enlargement. J Am Soc Hortic Sci 138:443–451Lloret A, Conejero A, Leida C et al (2017) Dual regulation of water retention and cell growth by a stress-associated protein (SAP) gene in Prunus. Sci Rep 7:332. doi: 10.1038/s41598-017-00471-7Lo Bianco R, Rieger M, Sung S-JS (2000) Effect of drought on sorbitol and sucrose metabolism in sinks and sources of peach. Physiol Plant 108:71–78. doi: 10.1034/j.1399-3054.2000.108001071.xLoescher WH (1987) Physiology and metabolism of sugar alcohols in higher-plants. Physiol Plant 70:553–557. doi: 10.1111/j.1399-3054.1987.tb02857.xLoescher WH, Marlow GC, Kennedy RA (1982) Sorbitol metabolism and sink-source interconversions in developing apple leaves. Plant Physiol 70:335–339. doi: 10.1104/pp.70.2.335Marquat C, Vandamme M, Gendraud M, Pétel G (1999) Dormancy in vegetative buds of peach: relation between carbohydrate absorption potentials and carbohydrate concentration in the bud during dormancy and its release. Sci Hortic 79:151–162. doi: 10.1016/S0304-4238(98)00203-9Niu Q, Li J, Cai D et al (2016) Dormancy-associated MADS-box genes and microRNAs jointly control dormancy transition in pear (Pyrus pyrifolia white pear group) flower bud. J Exp Bot 67:239–257. doi: 10.1093/jxb/erv454Pfaffl MW, Tichopad A, Prgomet C, Neuvians TP (2004) Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper—excel-based tool using pair-wise correlations. Biotechnol Lett 26:509–515. doi: 10.1023/B:BILE.0000019559.84305.47Ríos G, Leida C, Conejero A, Badenes ML (2014) Epigenetic regulation of bud dormancy events in perennial plants. Front Plant Sci 5:247. doi: 10.3389/fpls.2014.00247Saito T, Bai S, Imai T, Ito A, Nakajima I, Moriguchi T (2015) Histone modification and signalling cascade of the dormancy-associated MADS-box gene, PpMADS13-1, in Japanese pear (Pyrus pyrifolia) during endodormancy. Plant Cell Environ 38:1157–1166. doi: 10.1111/pce.12469Santamaría ME, Hasbún R, Valera MJ et al (2009) Acetylated H4 histone and genomic DNA methylation patterns during bud set and bud burst in Castanea sativa. J Plant Physiol 166:1360–1369. doi: 10.1016/j.jplph.2009.02.014Shen B, Hohmann S, Jensen RG, Bohnert HJ (1999) Roles of sugar alcohols in osmotic stress adaptation replacement of glycerol by mannitol and sorbitol in yeast. Plant Physiol 121:45–52. doi: 10.1104/pp.121.1.45Sheveleva EV, Marquez S, Chmara W, Zegeer A, Jensen RG, Bohnert HJ (1998) Sorbitol-6-phosphate dehydrogenase expression in transgenic tobacco high amounts of sorbitol lead to necrotic lesions. Plant Physiol 117:831–839. doi: 10.1104/pp.117.3.831Silver N, Best S, Jian J, Thein SL (2006) Selection of housekeeping genes for gene expression studies in human reticulocytes using real-time PCR. BMC Mol Biol 7:33. doi: 10.1186/1471-2199-7-33Talavera G, Castresana J (2007) Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Syst Biol 56:564–577. doi: 10.1080/10635150701472164Tao R, Uratsu SL, Dandekar AM (1995) Sorbitol synthesis in transgenic tobacco with apple cDNA encoding NADP-dependent sorbitol-6-phosphate dehydrogenase. Plant Cell Physiol 36:525–532. doi: 10.1093/oxfordjournals.pcp.a078789Teo G, Suzuki Y, Uratsu SL et al (2006) Silencing leaf sorbitol synthesis alters long-distance partitioning and apple fruit quality. Proc Natl Acad Sci USA 103:18842–18847. doi: 10.1073/pnas.0605873103Trotel P, Bouchereau A, Niogret MF, Larher F (1996) The fate of osmo-accumulated proline in leaf discs of Rape (Brassica napus L) incubated in a medium of low osmolarity. Plant Sci 118:31–45. doi: 10.1016/0168-9452(96)04422-6Verde I, Abbott AG, Scalabrin S et al (2013) The high-quality draft genome of peach (Prunus persica) identifies unique patterns of genetic diversity, domestication and genome evolution. Nat Genet 45:487–494. doi: 10.1038/ng.2586Webb KL, Burley JWA (1962) Sorbitol translocation in apple. Science 137:766. doi: 10.1126/science.137.3532.766Wisniewski M, Norelli J, Artlip T (2015) Overexpression of a peach CBF gene in apple: a model for understanding the integration of growth, dormancy, and cold hardiness in woody plants. Front Plant Sci 6:85. doi: 10.3389/fpls.2015.00085Yadav R, Prasad R (2014) Identification and functional characterization of sorbitol-6-phosphate dehydrogenase protein from rice and structural elucidation by in silico approach. Planta 240:223–238. doi: 10.1007/s00425-014-2076-

Chilling-Dependent Release of Seed and Bud Dormancy in Peach Associates to Common Changes in Gene Expression

Reproductive meristems and embryos display dormancy mechanisms in specialized structures named respectively buds and seeds that arrest the growth of perennial plants until environmental conditions are optimal for survival. Dormancy shows common physiological features in buds and seeds. A genotype-specific period of chilling is usually required to release dormancy by molecular mechanisms that are still poorly understood. In order to find common transcriptional pathways associated to dormancy release, we analyzed the chilling-dependent expression in embryos of certain genes that were previously found related to dormancy in flower buds of peach. We propose the presence of short and long-term dormancy events affecting respectively the germination rate and seedling development by independent mechanisms. Short periods of chilling seem to improve germination in an abscisic acid-dependent manner, whereas the positive effect of longer cold treatments on physiological dwarfing coincides with the accumulation of phenylpropanoids in the seed

Additive QTLs on three chromosomes control flowering time in woodland strawberry (Fragaria vesca L.)

Flowering time is an important trait that affects survival, reproduction and yield in both wild and cultivated plants. Therefore, many studies have focused on the identification of flowering time quantitative trait locus (QTLs) in different crops, and molecular control of this trait has been extensively investigated in model species. Here we report the mapping of QTLs for flowering time and vegetative traits in a large woodland strawberry mapping population that was phenotyped both under field conditions and in a greenhouse after flower induction in the field. The greenhouse experiment revealed additive QTLs in three linkage groups (LG), two on both LG4 and LG7, and one on LG6 that explain about half of the flowering time variance in the population. Three of the QTLs were newly identified in this study, and one co-localized with the previously characterized FvTFL1 gene. An additional strong QTL corresponding to previously mapped PFRU was detected in both field and greenhouse experiments indicating that gene(s) in this locus can control the timing of flowering in different environments in addition to the duration of flowering and axillary bud differentiation to runners and branch crowns. Several putative flowering time genes were identified in these QTL regions that await functional validation. Our results indicate that a few major QTLs may control flowering time and axillary bud differentiation in strawberries. We suggest that the identification of causal genes in the diploid strawberry may enable fine tuning of flowering time and vegetative growth in the closely related octoploid cultivated strawberry.Peer reviewe

Control of Flowering in Strawberries

Strawberries (Fragaria sp.) are small perennial plants capable of both sexual reproduction through seeds and clonal reproduction via runners. Because vegetative and generative developmental programs are tightly connected, the control of flowering is presented here in the context of the yearly growth cycle. The rosette crown of strawberry consists of a stem with short internodes produced from the apical meristem. Each node harbors one trifoliate leaf and an axillary bud. The fate of axillary buds is dictated by environmental conditions; high temperatures and long days (LDs) promote axillary bud development into runners, whereas cool temperature and short days (SDs) favor the formation of branch crowns. SDs and cool temperature also promote flowering; under these conditions, the main shoot apical meristem is converted into a terminal inflorescence, and vegetative growth is continued from the uppermost axillary branch crown. The environmental factors that regulate vegetative and generative development in strawberries have been reasonably well characterized and are reviewed in the first two chapters. The genetic basis of the physiological responses in strawberries is much less clear. To provide a point of reference for the flowering pathways described in strawberries so far, a short review on the molecular mechanisms controlling flowering in the model plant Arabidopsis is given. The last two chapters will then describe the current knowledge on the molecular mechanisms controlling the physiological responses in strawberries.Peer reviewe

Mapping of Candidate Genes Involved in Bud Dormancy and Flowering Time in Sweet Cherry (Prunus avium)

The timing of flowering in perennial plants is crucial for their survival in temperate climates and is regulated by the duration of bud dormancy. Bud dormancy release and bud break depend on the perception of cumulative chilling during endodormancy and heat during the bud development. The objectives of this work were to identify candidate genes involved in dormancy and flowering processes in sweet cherry, their mapping in two mapping progenies 'Regina' × 'Garnet' and 'Regina' × 'Lapins', and to select those candidate genes which co-localized with quantitative trait loci (QTLs) associated with temperature requirements for bud dormancy release and flowering. Based on available data on flowering processes in various species, a list of 79 candidate genes was established. The peach and sweet cherry orthologs were identified and primers were designed to amplify sweet cherry candidate gene fragments. Based on the amplified sequences of the three parents of the mapping progenies, SNPs segregations in the progenies were identified. Thirty five candidate genes were genetically mapped in at least one of the two progenies and all were in silico mapped. Co-localization between candidate genes and QTLs associated with temperature requirements and flowering date were identified for the first time in sweet cherry. The allelic composition of the candidate genes located in the major QTL for heat requirements and flowering date located on linkage group 4 have a significant effect on these two traits indicating their potential use for breeding programs in sweet cherry to select new varieties adapted to putative future climatic conditions