HLA-G gene editing in tumor cell lines as a novel alternative in cancer immunotherapy

Cancer immunotherapies based mainly on the blockade of immune-checkpoint (IC) molecules by anti-IC antibodies offer new alternatives for treatment in oncological diseases. However, a considerable proportion of patients remain unresponsive to them. Hence, the development of novel clinical immunotherapeutic approaches and/or targets are crucial.W In this context, targeting the immune-checkpoint HLA-G/ILT2/ILT4 has caused great interest since it is abnormally expressed in several malignancies generating a tolerogenic microenvironment. Here, we used CRISPR/Cas9 gene editing to block the HLA-G expression in two tumor cell lines expressing HLA-G, including a renal cell carcinoma (RCC7) and a choriocarcinoma (JEG-3). Different sgRNA/Cas9 plasmids targeting HLA-G exon 1 and 2 were transfected in both cell lines. Downregulation of HLA-G was reached to different degrees, including complete silencing. Most importantly, HLA-G − cells triggered a higher in vitro response of immune cells with respect to HLA-G + wild type cells. Altogether, we demonstrated for the first time the HLA-G downregulation through gene editing. We propose this approach as a first step to develop novel clinical immunotherapeutic approaches in cancer.Fil: Palma, Maria Belen. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Departamento de Ciencias Morfológicas. Cátedra de Citología y Embriología A; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Tronik Le Roux, Diana. Saint-Louis Hospital; Francia. Universite de Paris 1 - Pantheon Sorbonne.; FranciaFil: Amin, Guadalupe. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Castañeda, Sheila Lucia. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Möbbs, Alan Miqueas. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Scarafia, Maria Agustina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: la Greca, Alejandro Damián. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Daouya, Marina. Hôpital Saint-louis; Francia. Universite de Paris 1 - Pantheon Sorbonne.; FranciaFil: Poras, Isabelle. Hôpital Saint-louis; Francia. Universite de Paris 1 - Pantheon Sorbonne.; FranciaFil: Inda, Ana María. Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Departamento de Ciencias Morfológicas. Cátedra de Citología y Embriología A; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; ArgentinaFil: Moro, Lucía Natalia. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Carosella, Edgardo Delfino. Hôpital Saint-louis; Francia. Universite de Paris 1 - Pantheon Sorbonne.; FranciaFil: García, Marcela Nilda. Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Departamento de Ciencias Morfológicas. Cátedra de Citología y Embriología A; ArgentinaFil: Miriuka, Santiago Gabriel. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Electrochemical impedance spectroscopy: a non-destructive method to study the corrosion of aluminum-magnesium alloys in a magnesium phosphate cement-based matrix

International audienc

Electrochemical impedance spectroscopy: a non-destructive method to study the corrosion of aluminum-magnesium alloys in a magnesium phosphate cement-based matrix

International audienc

Electrochemical Behavior of Al/Mg Alloys Immobilized in a Magnesium Potassium Phosphate Cement-Based Mortar

International audiencePortland cement is extensively used for the conditioning of radioactive waste. However, its high alkalinity is a serious obstacle to the stabilization of waste containing aluminum metal since aluminum is oxidized by the pore solution with the production of dihydrogen. This work investigates the potential of an alternative binder, magnesium potassium phosphate (MKP) cement, for the stabilization of Al–Mg alloys comprising 2 to 4.5 wt% of Mg and other metallic impurities. The objective is to assess the influence of the alloy composition on its reactivity in the cementitious matrix at earlier ages, as well as at later ages, when the cement has reached a significant reaction degree. Two complementary techniques are used. Gas chromatography shows that the dihydrogen release, resulting from the corrosion process, is not influenced by the magnesium content in the alloy. Electrochemical impedance spectroscopy provides qualitative information about the corrosion but also makes it possible to assess the corrosion current using an equivalent electrical circuit linked to the kinetic parameters of the postulated corrosion mechanism. Over a one-year period, the corrosion current of the alloys, regardless of their Mg content, is reduced by almost three orders of magnitude in MKP mortar as compared to Portland-cement-based mortar.</jats:p

Electrochemical impedance spectroscopy: a non-destructive method to study the corrosion of aluminum-magnesium alloys in a magnesium phosphate cement-based matrix

International audienc

Study of the conditioning of aluminum-magnesium alloys in magnesium potassium phosphate cement

International audienc

Magnesium potassium phosphate cement: a promising binder for the conditioning of aluminum-magnesium alloys waste

International audienceThe reprocessing of spent fuel designed for natural uranium – graphite – gas reactors has produced somewaste with aluminum alloys, which need to be stabilized and solidified before their final disposal. Portlandcement is extensively used for the conditioning of low-level and intermediate-level radioactive waste;however, its high alkalinity is a serious obstacle to aluminum stabilization, as it is oxidized by the mixingsolution, with production of dihydrogen. This work investigates a new solution consisting in usingmagnesium potassium phosphate cement (MKPC) instead of Portland cement (PC). Gas chromatographyand electrochemical impedance spectroscopy (EIS) are used to monitor the corrosion of pure aluminum and aluminum-magnesium alloys containing 2 to 4.5 wt.% of Mg in MKPC mortar. EIS provides qualitativeinformation about the corrosion, but also makes it possible to assess the corrosion current using anequivalent electrical circuit linked to the kinetic parameters of the postulated corrosion mechanism. It isshown that the corrosion current of the alloys, regardless of their composition, is reduced by about twoorders of magnitude in MKPC mortar as compared to Portland cement mortar. This result opens up newprospects for increasing the incorporation rate of reactive Al metal in a cementitious matrix

Magnesium potassium phosphate cement: a promising binder for the conditioning of aluminum-magnesium alloys waste

International audienceThe reprocessing of spent fuel designed for natural uranium – graphite – gas reactors has produced somewaste with aluminum alloys, which need to be stabilized and solidified before their final disposal. Portlandcement is extensively used for the conditioning of low-level and intermediate-level radioactive waste;however, its high alkalinity is a serious obstacle to aluminum stabilization, as it is oxidized by the mixingsolution, with production of dihydrogen. This work investigates a new solution consisting in usingmagnesium potassium phosphate cement (MKPC) instead of Portland cement (PC). Gas chromatographyand electrochemical impedance spectroscopy (EIS) are used to monitor the corrosion of pure aluminum and aluminum-magnesium alloys containing 2 to 4.5 wt.% of Mg in MKPC mortar. EIS provides qualitativeinformation about the corrosion, but also makes it possible to assess the corrosion current using anequivalent electrical circuit linked to the kinetic parameters of the postulated corrosion mechanism. It isshown that the corrosion current of the alloys, regardless of their composition, is reduced by about twoorders of magnitude in MKPC mortar as compared to Portland cement mortar. This result opens up newprospects for increasing the incorporation rate of reactive Al metal in a cementitious matrix

Contribution of electrochemical impedance spectroscopy to the study of aluminium-magnesium alloys corrosion in magnesium potassium phosphate cement pore solution

International audienc

Contribution of electrochemical impedance spectroscopy to the study of aluminium-magnesium alloys corrosion in magnesium potassium phosphate cement pore solution

International audienc