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morphology and size of the crystals (~40 nm), that are paramagnetic at room and body temperature. Because
of this protein, BMNPs have a negative surface charge at physiological pH values that allow an efficient
coupling with different molecules.

These BMNPs were functionalized with the chemotherapy drug doxorubicin (DOXO) and their ability
to respond to an externally applied gradient magnetic field (GMF) was studied both in vitro and in vivo.
Naïve BMNPs were cytocompatibles on 4T1 cells, while the DOXO-BMNPs were toxic starting from short
exposure times when a GMF is applied. This allowed a faster interaction between BMNPs and cells (Perls
Blue Staining) as well as a faster delivery of the DOXO to cell nuclei (confocal analysis). When DOXO-
functionalized or not functionalized BMNPs were i.v. injected in mice bearing 4T1 cells-induced tumors,
the application of the magnet on the tumors for 1 h reduced their size. Moreover, the presence of DOXO on
BMNPs enhanced this effect.

All together, these data suggest that tumor attack by combined strategies (chemotherapeutic drug and
magnetic field) could represent a promising approach for cancer therapy. Future steps will be the possibility
to apply an alternating magnetic field to BMNPs to induce hyperthermia, to which tumor cells are more
sensitive than healthy cells, and to enhance a more efficient localized release of the drug at the tumor site.

35. Novel diacylglycerol kinase alpha inhibitors for X-linked lymphoproliferative disease 1
therapy

1,3
4
5
Suresh Velnati , Elisa Ruffo , Alberto Massarotti , Maria Talmon , Sai Sandeep Varma
1,2
5
1,2
1
4
6
Konduru , Annamaria Antona , Alessandro Gesu , Luigia Grazia Fresu , Andrew L Snow ,
1
1
4
1,3
Daniela Capello , Alessandra Bertoni , Gian Cesare Tron , Andrea Graziani , Gianluca
Baldanzi 1,2
1 Department of Translational Medicine, University of Piemonte Orientale, via Solaroli 17, Novara 28100, Italy.
2 Center for translational research on Allergy and Autoimmune Diseases (CAAD), Novara 28100, Italy.
3 Division of Experimental Oncology, School of Medicine, University Vita e Salute San Raffaele, Milan 20132,
Italy.
4 Department of Pharmaceutical Sciences, University of Piemonte Orientale, largo Donegani 12, Novara 28100,
Italy.
5 Department of Health Sciences, School of Medicine, University of Piemonte Orientale, Novara 28100, Italy.
6 Department of Pharmacology and Molecular Therapeutics, Uniformed Services University of the Health
Sciences, Bethesda, MD 20814, USA.
Background: X-linked lymphoproliferative disease 1 (XLP1) is a primary immunodeficiency due to
mutations in the SH2D1a gene, encoding SAP. SAP deficiency perturbs TCR signalling and results in a
constitutive diacylglycerol kinase alpha (DGKα) activity that impairs CD8+ T cell restimulation induced
cell death (RICD). Indeed, pharmacological inhibition of DGKα in XLP1 animal models limits CD8+ T cell
expansion and interferon-γ production, suggesting the development of DGKα inhibitors for XLP1 therapy.

Experimental model: To find new DGKα inhibitors, we used a 2D/3D in silico approach based on chemical
homology with the two commercially available DGKα inhibitors (R59922 and R59949). The library was
screened for inhibitory activity at 100 µmol/L. Active compounds were tested at concentration from 0.1 to
100 µmol/L to estimate the IC . Furthermore, we tested the most active compounds in RICD assay using
50
SAP silenced lymphocytes as XLP1 model.