Suchergebnisse

[Purpose]: Mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL) remain B-cell malignancies with limited therapeutic options. The present study investigates the in vitro and in vivo effect of the phospholipid ether edelfosine (1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine) in MCL and CLL. [Experimental Design]: Several cell lines, patient-derived tumor cells, and xenografts in severe combined immunodeficient mice were used to examine the anti-MCL and anti-CLL activity of edelfosine. Furthermore, we analyzed the mechanism of action and drug biodistribution of edelfosine in MCL and CLL tumor-bearing severe combined immunodeficient mice. [Results]: Here, we have found that the phospholipid ether edelfosine was the most potent alkyl-lysophospholipid analogue in killing MCL and CLL cells, including patient-derived primary cells, while sparing normal resting lymphocytes. Alkyl-lysophospholipid analogues ranked edelfosine > perifosine ≫ erucylphosphocholine ≥ miltefosine in their capacity to elicit apoptosis in MCL and CLL cells. Edelfosine induced coclustering of Fas/CD95 death receptor and rafts in MCL and CLL cells. Edelfosine was taken up by malignant cells, whereas normal resting lymphocytes hardly incorporated the drug. Raft disruption by cholesterol depletion inhibited drug uptake, Fas/CD95 clustering, and edelfosine-induced apoptosis. Edelfosine oral administration showed a potent in vivo anticancer activity in MCL and CLL xenograft mouse models, and the drug accumulated dramatically and preferentially in the tumor. [Conclusions]: Our data indicate that edelfosine accumulates and kills MCL and CLL cells in a rather selective way, and set coclustering of Fas/CD95 and lipid rafts as a new framework in MCL and CLL therapy. Our data support a selective antitumor action of edelfosine. ; Ministerio de Ciencia e Innovación grants SAF2005-04293, SAF2006-8850, SAF2007-61261, SAF2007-60964, PCT-090100-2007-27, PS09/01915, and SAF2008-02251; Red Temática de Investigación Cooperativa en Cáncer grants RD06/0020/0014, RD06/0020/0097, and RD06/0020/1037 (Instituto de Salud Carlos III, cofunded by the Fondo Europeo de Desarrollo Regional of the European Union); Fondo de Investigación Sanitaria and European Commission grant FIS-FEDER 06/0813; Junta de Castilla y León (CSI01A08, GR15-Experimental Therapeutics and Translational Oncology Program, and Biomedicine Project 2009); Fundación de Investigación Médica Mutua Madrileña; Fundación "la Caixa" grant BM05-30-0; Caja Navarra Foundation; Department of Health of the Government of Navarra ("Ortiz de Landázuri, 2009" project); and AGAUR-Generalitat de Catalunya grant 2005SGR-00549. C. Gajate is supported by the Ramón y Cajal Program from the Ministerio de Ciencia e Innovación of Spain. A. Estella-Hermoso de Mendoza is supported by Department of Education of the Basque Government research grant BFI06.37. M. de Frias is a recipient of a fellowship from the AGAUR-Generalitat de Catalunya. G. Roué holds a Miguel Servet research contract from Instituto de Salud Carlos III. ; Peer Reviewed

15 Págs. ; Several psychiatric, neurologic and neurodegenerative disorders present increased brain ventricles volume, being hydrocephalus the disease with the major manifestation of ventriculomegaly caused by the accumulation of high amounts of cerebrospinal fluid (CSF). The molecules and pathomechanisms underlying cerebral ventricular enlargement are widely unknown. Kinase D interacting substrate of 220 kDa (KIDINS220) gene has been recently associated with schizophrenia and with a novel syndrome characterized by spastic paraplegia, intellectual disability, nystagmus and obesity (SINO syndrome), diseases frequently occurring with ventriculomegaly. Here we show that Kidins220, a transmembrane protein effector of various key neuronal signalling pathways, is a critical regulator of CSF homeostasis. We observe that both KIDINS220 and the water channel aquaporin-4 (AQP4) are markedly downregulated at the ventricular ependymal lining of idiopathic normal pressure hydrocephalus (iNPH) patients. We also find that Kidins220 deficient mice develop ventriculomegaly accompanied by water dyshomeostasis and loss of AQP4 in the brain ventricular ependymal layer and astrocytes. Kidins220 is a known cargo of the SNX27-retromer, a complex that redirects endocytosed plasma membrane proteins (cargos) back to the cell surface, thus avoiding their targeting to lysosomes for degradation. Mechanistically, we show that AQP4 is a novel cargo of the SNX27-retromer and that Kidins220 deficiency promotes a striking and unexpected downregulation of the SNX27-retromer that results in AQP4 lysosomal degradation. Accordingly, SNX27 silencing decreases AQP4 levels in wild-type astrocytes whereas SNX27 overexpression restores AQP4 content in Kidins220 deficient astrocytes. Together our data suggest that the KIDINS220-SNX27-retromer-AQP4 pathway is involved in human ventriculomegaly and open novel therapeutic perspectives. ; This work was supported by Spanish grants SAF2017-88885-R to TI and SAF2017-88881-R to MRC from the Spanish Ministerio de Economía y Competitividad/Agencia Estatal de Investigación/European Union Funds for Regional Development (MINECO/AEI/ FEDER, EU); by Comunidad de Madrid through the European Social Fund (ESF)-financed programme NEUROMETAB-CM (B2017/BMD-3700) to TI and AORTASANA-CM (B2017/BMD-3676) to MRC; by 'La Caixa' Banking Foundation project HR18-00068 to MRC; by Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED, Instituto de Salud Carlos III, Spain) CB06/05/1122 to TI and collaborative grant CIBERNED PI2018/06 to TI and IFa; by grant PID2019-104763RB-100 and Ramón y Cajal programme contract (RYC-2014-15991), both from MINECO/AEI/FEDER (EU) to EP; by Wellcome Trust Senior Investigator Award (107116/Z/15/Z), the European Union's Horizon 2020 Research and Innovation programme under grant agreement 739572 and a UK Dementia Research Institute Foundation award, all to GS; by grant PI19/00778 to AJJ from the Spanish Instituto de Salud Carlos III, co-funded by FEDER (EU); by Spanish grants from FEDER/Ministerio de Ciencia e Innovación (MICI)–AEI (SAF2017-86690-R), Network of Excellence iDIFFER (RED2018-102723-T), Instituto de Salud Carlos III (CIBERNED CB06/05/0086 and RETIC Tercel RD16/0011/0017) and Generalitat Valenciana (Prometeo 2017/030) to IFa. ADP was funded by a Juan de la Cierva Formación programme contract (FJCI-2014-19673; MINECO) and CIBERNED; JP-U was contracted by SAF2017-88885-R grant; AS-S was contracted by grants CIBERNED-PI2015-2/06 and PI2018/06, and SAF2017-88885-R; AS-G was funded by a contract from B2017/BMD-3700 grant and LSMP by a PhD training contract (PRE2018-086989) associated to grant SAF2017-88885-R. The cost of this publication has been paid in part by FEDER (European Union Funds for Regional Development) funds. We are particularly indebted to the patients and the Biobanks integrated in the Spanish National Biobanks Network, Biobank Banco de Tejidos CIEN (PT17/0015/0014), the HCB-IDIBAPS Biobank and Institute of Neuropathology Brain Bank-IDIBELL, for their collaboration in sample and data procurement. We are grateful to Professor M. Amiry-Moghaddam (University of Oslo, Norway) for his help with AQP4 reagents and methodological details, Professors P.J. Cullen (University of Bristol, UK) and F. Steinberg (Albert Ludwigs Universitaet Freiburg, Germany) for providing SNX27 plasmids. We also acknowledge M. Rodríguez-Martínez and T. Navarro for performing MRI studies; Drs. P. Lopez-Larrubia (IIBM, CSIC-UAM) and A. Martín (Achucarro Basque Center for Neuroscience, Bilbao, Spain) for advice in MRI analysis; L. García-Guerra, A. González-Martín and other laboratory members for help and suggestions; L. Sánchez-Ruiloba for technical assistance in confocal images analysis, L. Jiménez for taking care of mice colonies; and to core facilities at Instituto de Investigaciones Biomédicas 'Alberto Sols' de Madrid (IIBM, CSIC-UAM): Animal Care, Genomics, Confocal microscopy. ; Peer reviewed

Several psychiatric, neurologic and neurodegenerative disorders present increased brain ventricles volume, being hydrocephalus the disease with the major manifestation of ventriculomegaly caused by the accumulation of high amounts of cerebrospinal fluid (CSF). The molecules and pathomechanisms underlying cerebral ventricular enlargement are widely unknown. Kinase D interacting substrate of 220 kDa (KIDINS220) gene has been recently associated with schizophrenia and with a novel syndrome characterized by spastic paraplegia, intellectual disability, nystagmus and obesity (SINO syndrome), diseases frequently occurring with ventriculomegaly. Here we show that Kidins220, a transmembrane protein effector of various key neuronal signalling pathways, is a critical regulator of CSF homeostasis. We observe that both KIDINS220 and the water channel aquaporin-4 (AQP4) are markedly downregulated at the ventricular ependymal lining of idiopathic normal pressure hydrocephalus (iNPH) patients. We also find that Kidins220 deficient mice develop ventriculomegaly accompanied by water dyshomeostasis and loss of AQP4 in the brain ventricular ependymal layer and astrocytes. Kidins220 is a known cargo of the SNX27-retromer, a complex that redirects endocytosed plasma membrane proteins (cargos) back to the cell surface, thus avoiding their targeting to lysosomes for degradation. Mechanistically, we show that AQP4 is a novel cargo of the SNX27-retromer and that Kidins220 deficiency promotes a striking and unexpected downregulation of the SNX27-retromer that results in AQP4 lysosomal degradation. Accordingly, SNX27 silencing decreases AQP4 levels in wild-type astrocytes whereas SNX27 overexpression restores AQP4 content in Kidins220 deficient astrocytes. Together our data suggest that the KIDINS220-SNX27-retromer-AQP4 pathway is involved in human ventriculomegaly and open novel therapeutic perspectives. ; This work was supported by Spanish grants SAF2017-88885-R to TI and SAF2017-88881-R to MRC from the Spanish Ministerio de Economía y Competitividad/Agencia Estatal de Investigación/European Union Funds for Regional Development (MINECO/AEI/ FEDER, EU); by Comunidad de Madrid through the European Social Fund (ESF)-financed programme NEUROMETAB-CM (B2017/BMD-3700) to TI and AORTASANA-CM (B2017/BMD-3676) to MRC; by 'La Caixa' Banking Foundation project HR18-00068 to MRC; by Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED, Instituto de Salud Carlos III, Spain) CB06/05/1122 to TI and collaborative grant CIBERNED PI2018/06 to TI and IFa; by grant PID2019-104763RB-100 and Ramón y Cajal programme contract (RYC-2014-15991), both from MINECO/AEI/FEDER (EU) to EP; by Wellcome Trust Senior Investigator Award (107116/Z/15/Z), the European Union's Horizon 2020 Research and Innovation programme under grant agreement 739572 and a UK Dementia Research Institute Foundation award, all to GS; by grant PI19/00778 to AJJ from the Spanish Instituto de Salud Carlos III, co-funded by FEDER (EU); by Spanish grants from FEDER/Ministerio de Ciencia e Innovación (MICI)–AEI (SAF2017-86690-R), Network of Excellence iDIFFER (RED2018-102723-T), Instituto de Salud Carlos III (CIBERNED CB06/05/0086 and RETIC Tercel RD16/0011/0017) and Generalitat Valenciana (Prometeo 2017/030) to IFa. ADP was funded by a Juan de la Cierva Formación programme contract (FJCI-2014-19673; MINECO) and CIBERNED; JP-U was contracted by SAF2017-88885-R grant; AS-S was contracted by grants CIBERNED-PI2015-2/06 and PI2018/06, and SAF2017-88885-R; AS-G was funded by a contract from B2017/BMD-3700 grant and LSMP by a PhD training contract (PRE2018-086989) associated to grant SAF2017-88885-R.