Adenovirus-transduced CD34 IntroductionGenetically modified stem/progenitor cells represent an innovative approach for delivery of anticancer molecules. 1,2 Because of their homing properties, systemically injected stem/progenitor cells could infiltrate both primary and metastatic tumor sites, thus allowing tumor-specific targeting 3-9 and potentially overcoming limitations inherent to the pharmacokinetic profile of soluble drugs. [10][11][12] Soluble tumor necrosis factor (TNF)-related apoptosisinducing ligand (sTRAIL) is a proapoptotic member of the TNF superfamily of death receptor ligands. A variety of preclinical data show that sTRAIL is a cancer cell-specific molecule exerting a remarkable antitumor activity in vitro [13][14][15][16][17][18] and in vivo in athymic nude mice or in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. 13,19,20 Phase 1/2 clinical trials have demonstrated a good toxicity profile for sTRAIL but limited evidence of antitumor activity probably because of short exposure of tumor cells to low drug concentrations. 21 Because of sTRAIL's short half-life, 13,20,22 it seems unlikely that the recommended sTRAIL dose of 8 mg/kg body weight will allow prolonged exposure of tumor cells at high drug concentrations. 21 Strategies to enhance the therapeutic activity of sTRAIL include combining it with conventional chemotherapy 23 or with new agents, such as histone deacetylase inhibitors that up-regulate TRAIL-R1 and/or TRAIL-R2, resulting in a synergistic induction of apoptosis in both sTRAIL-sensitive and -resistant tumor cells. 24 Alternatively, cell-based vehiculation of the full-length, membranebound (m)TRAIL has been proposed. 11 Neural or mesenchymal stem cell-mediated mTRAIL delivery has been investigated in solid tumors. 9,[25][26][27][28] More recently, we demonstrated that intravenous injection of mTRAIL-expressing CD34 ϩ cells (CD34-TRAIL ϩ cells) efficiently act as mTRAIL-presenting vehicles and exert a potent antitumor activity in NOD/SCID mice bearing systemic multiple myeloma and non-Hodgkin lymphoma xenografts. [29][30][31] Using a subcutaneous multiple myeloma model in NOD/SCID mice, the present study aimed at investigating the antitumor mechanism(s) of mTRAIL-expressing cells by analyzing homing properties of CD34-TRAIL ϩ cells as well as the degree and distribution of tumor cell death and tumor vascular damage. To analyze the antivascular activity of CD34-TRAIL ϩ cells, we used an in vivo staining assay of tumor vasculature. 32 The imaging software ImageJ (National Institutes of Health), equipped with appropriately designed macros to specifically quantify parameters of tumor cell and tumor vascular damage, allowed the processing The online version of this article contains a data supplement.The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked ''advertisement'' in accordance with 18 USC section 1734. For personal use only. on May 11, 2018. by guest www.blood...

In regenerative medicine, human cord blood-derived multipotent mesenchymal stromal cells (CBMSCs) stand out for their biological peculiarities demonstrated in in vitro and in vivo preclinical studies. Here, we present our 9-year experience for the consistent isolation of CBMSCs. Although nearly one CB unit out of two retains the potential to give rise to MSC colonies, only 46% of them can be cultured till low passages (P ‡ 4), but onefourth of those reaches even higher passages (P ‡ 8). Subsequent characterization for morphological, clonal, differentiation, and proliferation properties revealed two divergent CBMSC behaviors. In particular, a cumulative population doublings cut-off (CPD = 15) was identified that undoubtedly distinguishes two growth curves, and different degrees of commitment toward osteogenesis were observed. These data clearly show the existence of at least two distinct CBMSC subsets: one mainly short-living and less proliferative (SL-CBMSCs), the other long-living, with higher growth rate, and, very importantly, with significantly (P £ 0.01) longer telomere (LLCBMSCs). Moreover, significant differences in the immunoprofile before seeding were found among CB units giving rise to LL-CBMSCs or SL-CBMSCs or showing no colony formation. Finally, all the aforementioned results provided a peculiar and useful set of parameters potentially predictive for CBMSC culture outcome.

BackgroundThe trophic, anti-apoptotic and regenerative effects of bone marrow mesenchymal stromal cells (MSC) may reduce neuronal cell loss in neurodegenerative disorders.MethodsWe used MSC as a novel candidate therapeutic tool in a pilot phase-I study for patients affected by progressive supranuclear palsy (PSP), a rare, severe and no-option form of Parkinsonism. Five patients received the cells by infusion into the cerebral arteries. Effects were assessed using the best available motor function rating scales (UPDRS, Hoehn and Yahr, PSP rating scale), as well as neuropsychological assessments, gait analysis and brain imaging before and after cell administration.ResultsOne year after cell infusion, all treated patients were alive, except one, who died 9 months after the infusion for reasons not related to cell administration or to disease progression (accidental fall). In all treated patients motor function rating scales remained stable for at least six-months during the one-year follow-up.ConclusionsWe have demonstrated for the first time that MSC administration is feasible in subjects with PSP. In these patients, in whom deterioration of motor function is invariably rapid, we recorded clinical stabilization for at least 6 months. These encouraging results pave the way to the next randomized, placebo-controlled phase-II study that will definitively provide information on the efficacy of this innovative approach.Trial registration ClinicalTrials.gov NCT01824121

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Platelet gel derived from peripheral blood is widely applied in many clinical fields of surgery as biomaterial containing growth factors with high proliferative properties. In 2010, we studied and patented a platelet gel derived from cord blood. In this study, due to the crucial role of the factors released by the platelet gel, we first extended the characterization of its releasate. Using a wide proteomic array and splitting the two components of the releasate, that is, platelets and plasma, we have been able to study their growth factor content. Interestingly, we discovered high levels of hormones and molecules able to support tissue growth in the cord blood platelet gel releasate and, in addition, higher concentrations of several angiogenic factors if compared with the peripheral blood counterpart. On the contrary, the latter was much richer in inflammatory factors. The second aim of our work was to study the effects on cell culture, immunophenotype, and function of mesenchymal stem cells exposed to these two platelet gel releasates as substitute for the animal serum. Since our findings nicely show that the use of the peripheral versus the cord blood platelet gel releasate can differently influence the mesenchymal stem cell commitment, we can suggest that in addition to its peculiar angiogenic properties cord blood platelet gel releasate shows excellent proliferative properties as cell culture supplement.

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The fully human anti-HLA-DR antibody 1D09C3 has been shown to delay lymphoma cell growth in severe combined immunodeficient (SCID) mice. The present study was aimed at (a) investigating the mechanism(s) of 1D09C3-induced cell death and (b) further exploring the therapeutic efficacy of 1D09C3 in nonobese diabetic (NOD)/SCID mice. The chronic lymphocytic leukemia cell line JVM-2 and the mantle cell lymphoma cell line GRANTA-519 were used. Generation of reactive oxygen species (ROS) and mitochondrial membrane depolarization were measured by flow cytometry following cell incubation with dihydroethidium and TMRE, respectively. Western blot analysis was used to detect c-Jun-NH 2 -kinase (JNK) phosphorylation and apoptosis-inducing factor (AIF). NOD/SCID mice were used to investigate the activity of 1D09C3 in early-or advanced-stage tumor xenografts. In vitro, 1D09C3-induced cell death involves a cascade of events, including ROS increase, JNK activation, mitochondrial membrane depolarization, and AIF release from mitochondria. Inhibition of JNK activity significantly reduced 1D09C3-induced apoptosis, indicating that 1D09C3 activity involves activation of the kinase. In vivo, 1D09C3 induces long-term disease-free survival in a significant proportion of tumorbearing mice treated at an early stage of disease. Treatment of mice bearing advanced-stage lymphoma results in a highly significant prolongation of survival. These data show that 1D09C3 (a) exerts a potent antitumor effect by activating ROSdependent, JNK-driven cell death, (b) cures the great majority of mice treated at an early-stage of disease, and (c) significantly prolongs survival of mice with advanced-stage disease. (Cancer Res 2006; 66(3): 1799-808)