Publications by authors named "Domenico Orlando"

7 Publications

  • Page 1 of 1

CD28.OX40 co-stimulatory combination is associated with long in vivo persistence and high activity of CAR.CD30 T-cells.

Haematologica 2021 04 1;106(4):987-999. Epub 2021 Apr 1.

Dept Onco-Haematology, Cell and Gene Therapy, Bambino Gesù Children Hospital, Rome, Italy.

The prognosis of many patients with chemotherapy-refractory or multiply relapsed CD30+ non-Hodgkin Lymphoma (NHL) or Hodgkin lymphoma (HL) still remains poor, and novel therapeutic approaches are warranted to address this unmet clinical need. In light of this consideration, we designed and pre-clinically validated a Chimeric Antigen Receptor (CAR) construct characterized by a novel anti-CD30 single-chain variable-fragment cassette, linked to CD3ζ by the signaling domains of two costimulatory molecules, namely either CD28.4-1BB or CD28.OX40. We found that CAR.CD30 T-cells exhibit remarkable cytolytic activity in vitro against HL and NHL cell lines, with sustained proliferation and pro-inflammatory cytokine production, even after multiple and sequential lymphoma cell challenges. CAR.CD30 T-cells also demonstrated anti-lymphoma activity in two in vivo xenograft immune-deficient mouse models of metastatic HL and NHL. We observed that administration of CAR.CD30 T-cells, incorporating the CD28.OX40 costimulatory domains and manufactured in the presence of IL7 and IL15, were associated with the best overall survival in the treated mice, along with the establishment of a long-term immunological memory, able to protect mice from further tumor re-challenge. Our data indicate that, in the context of in vivo systemic metastatic xenograft mouse models, the costimulatory machinery of CD28.OX40 is crucial for improving persistence, in vivo expansion and proliferation of CAR.CD30 T-cells upon tumor encounter. CD28.OX40 costimulatory combination is ultimately responsible for the antitumor efficacy of the approach, paving the way to translate this therapeutic strategy in patients with CD30+ HL and NHL.
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http://dx.doi.org/10.3324/haematol.2019.231183DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8018158PMC
April 2021

Conformal Dimensions in the Large Charge Sectors at the O(4) Wilson-Fisher Fixed Point.

Phys Rev Lett 2019 Aug;123(5):051603

Albert Einstein Center for Fundamental Physics, Institute for Theoretical Physics, University of Bern, Sidlerstrasse 5, ch-3012, Bern, Switzerland.

We study the O(4) Wilson-Fisher fixed point in 2+1 dimensions in fixed large-charge sectors identified by products of two spin-j representations (j_{L},j_{R}). Using effective field theory we derive a formula for the conformal dimensions D(j_{L},j_{R}) of the leading operator in terms of two constants, c_{3/2} and c_{1/2}, when the sum j_{L}+j_{R} is much larger than the difference |j_{L}-j_{R}|. We compute D(j_{L},j_{R}) when j_{L}=j_{R} with Monte Carlo calculations in a discrete formulation of the O(4) lattice field theory, and show excellent agreement with the predicted formula and estimate c_{3/2}=1.068(4) and c_{1/2}=0.083(3).
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http://dx.doi.org/10.1103/PhysRevLett.123.051603DOI Listing
August 2019

Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma.

Oncoimmunology 2018;7(6):e1433518. Epub 2018 Mar 15.

Department of Pediatric Hematology and Oncology, IRCCS Ospedale Pediatrico Bambino Gesù, Rome, Italy.

Chimeric antigen receptor (CAR) T-cell therapy has been shown to be dramatically effective in the treatment of B-cell malignancies. However, there are still substantial obstacles to overcome, before similar responses can be achieved in patients with solid tumors. We evaluated both and in a preclinical murine model the efficacy of different 2nd and 3rd generation CAR constructs targeting GD2, a disial-ganglioside expressed on the surface of neuroblastoma (NB) tumor cells. In order to address potential safety concerns regarding clinical application, an inducible safety switch, namely inducible Caspase-9 (iC9), was also included in the vector constructs. Our data indicate that a 3rd generation CAR incorporating CD28.4-1BB costimulatory domains is associated with improved anti-tumor efficacy as compared with a CAR incorporating the combination of CD28.OX40 domains. We demonstrate that the choice of 4-1BB signaling results into significant amelioration of several CAR T-cell characteristics, including: 1) T-cell exhaustion, 2) basal T-cell activation, 3) tumor control and 4) T-cell persistence. The fine-tuning of T-cell culture conditions obtained using IL7 and IL15 was found to be synergic with the CAR.GD2 design in increasing the anti-tumor activity of CAR T cells. We also demonstrate that activation of the suicide gene iC9, included in our construct without significantly impairing neither CAR expression nor anti-tumor activity, leads to a prompt induction of apoptosis of GD2.CAR T cells. Altogether, these findings are instrumental in optimizing the function of CAR T-cell products to be employed in the treatment of children with NB.
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http://dx.doi.org/10.1080/2162402X.2018.1433518DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5980417PMC
March 2018

Adoptive Immunotherapy Using PRAME-Specific T Cells in Medulloblastoma.

Cancer Res 2018 06 3;78(12):3337-3349. Epub 2018 Apr 3.

Department of Pediatric Haematology and Oncology, IRCCS Ospedale Pediatrico Bambino Gesù, Rome, Italy.

Medulloblastoma is the most frequent malignant childhood brain tumor with a high morbidity. Identification of new therapeutic targets would be instrumental in improving patient outcomes. We evaluated the expression of the tumor-associated antigen PRAME in biopsies from 60 patients with medulloblastoma. PRAME expression was detectable in 82% of tissues independent of molecular and histopathologic subgroups. High PRAME expression also correlated with worse overall survival. We next investigated the relevance of PRAME as a target for immunotherapy. Medulloblastoma cells were targeted using genetically modified T cells with a PRAME-specific TCR (SLL TCR T cells). SLL TCR T cells efficiently killed medulloblastoma HLA-A*02 DAOY cells as well as primary HLA-A*02 medulloblastoma cells. Moreover, SLL TCR T cells controlled tumor growth in an orthotopic mouse model of medulloblastoma. To prevent unexpected T-cell-related toxicity, an inducible caspase-9 (iC9) gene was introduced in frame with the SLL TCR; this safety switch triggered prompt elimination of genetically modified T cells. Altogether, these data indicate that T cells genetically modified with a high-affinity, PRAME-specific TCR and iC9 may represent a promising innovative approach for treating patients with HLA-A*02 medulloblastoma. These findings identify PRAME as a medulloblastoma tumor-associated antigen that can be targeted using genetically modified T cells. .
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http://dx.doi.org/10.1158/0008-5472.CAN-17-3140DOI Listing
June 2018

Conformal Dimensions via Large Charge Expansion.

Phys Rev Lett 2018 Feb;120(6):061603

Albert Einstein Center for Fundamental Physics, Institute for Theoretical Physics, University of Bern, Sidlerstrasse 5, CH-3012 Bern, Switzerland.

We construct an efficient Monte Carlo algorithm that overcomes the severe signal-to-noise ratio problems and helps us to accurately compute the conformal dimensions of large-Q fields at the Wilson-Fisher fixed point in the O(2) universality class. Using it, we verify a recent proposal that conformal dimensions of strongly coupled conformal field theories with a global U(1) charge can be obtained via a series expansion in the inverse charge 1/Q. We find that the conformal dimensions of the lowest operator with a fixed charge Q are almost entirely determined by the first few terms in the series.
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http://dx.doi.org/10.1103/PhysRevLett.120.061603DOI Listing
February 2018

Evaluation of Endoglin (CD105) expression in pediatric rhabdomyosarcoma.

BMC Cancer 2018 01 5;18(1):31. Epub 2018 Jan 5.

Department of Hematology/Oncology, Bambino Gesù Children's Hospital, IRCCS, Piazza di Sant'Onofrio, 4, 00165, Rome, Italy.

Background: The Intratumoral Microvessel Density (IMVD) is commonly used to quantify tumoral vascularization and is usually assessed by pan-endothelial markers, such as CD31. Endoglin (CD105) is a protein predominantly expressed in proliferating endothelium and the IMVD determined by this marker measures specifically the neovascularization. In this study, we investigated the CD105 expression in pediatric rhabdomyosarcoma and assessed the neovascularization by using the angiogenic ratio IMVD-CD105 to IMVD-CD31.

Methods: Paraffin-embedded archival tumor specimens were selected from 65 pediatric patients affected by rhabdomyosarcoma. The expression levels of CD105, CD31 and Vascular Endothelial Growth Factor (VEGF) were investigated in 30 cases (18 embryonal and 12 alveolar) available for this study. The IMVD-CD105 to IMVD-CD31 expression ratio was correlated with clinical and pathologic features of these patients.

Results: We found a specific expression of endoglin (CD105) in endothelial cells of all the rhabdomyosarcoma specimens analyzed. We observed a significant positive correlation between the IMVD individually measured by CD105 and CD31. The CD105/CD31 expression ratio was significantly higher in patients with lower survival and embryonal histology. Indeed, patients with a CD105/CD31 expression ratio < 1.3 had a significantly increased OS (88%, 95%CI, 60%-97%) compared to patients with higher values (40%, 95%CI, 12%-67%). We did not find any statistical correlation among VEGF and EFS, OS and CD105/CD31 expression ratio.

Conclusion: CD105 is expressed on endothelial cells of rhabdomyosarcoma and represent a useful tool to quantify neovascularization in this tumor. If confirmed by further studies, these results will indicate that CD105 is a potential target for combined therapies in rhabdomyosarcoma.
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http://dx.doi.org/10.1186/s12885-017-3947-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5755407PMC
January 2018

Inducible CTCF insulator delays the 3' regulatory region-mediated activation of germline promoters and alters class switching.

Proc Natl Acad Sci U S A 2017 06 22;114(23):6092-6097. Epub 2017 May 22.

Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, Université Paul Sabatier (UPS), 31077 Toulouse, France;

Class switch recombination (CSR) plays an important role in adaptive immune response by enabling mature B cells to switch from IgM expression to the expression of downstream isotypes. CSR is preceded by inducible germline (GL) transcription of the constant genes and is controlled by the 3' regulatory region (3'RR) in a stimulus-dependent manner. Why the 3'RR-mediated up-regulation of GL transcription is delayed to the mature B-cell stage is presently unknown. Here we show that mice devoid of an inducible CTCF binding element, located in the constant gene, display a marked isotype-specific increase of GL transcription in developing and resting splenic B cells and altered CSR in activated B cells. Moreover, insertion of a GL promoter downstream of the CTCF insulator led to premature activation of the ectopic promoter. This study provides functional evidence that the 3'RR has a developmentally controlled potential to constitutively activate GL promoters but that this activity is delayed, at least in part, by the CTCF insulator, which borders a transcriptionally active domain established by the 3'RR in developing B cells.
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http://dx.doi.org/10.1073/pnas.1701631114DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5468671PMC
June 2017