Immunomodulatory oligonucleotide IMT504: effects on mesenchymal stem cells as a first-in-class immunoprotective/immunoregenerative therapy

The immune responses of humans and animals to insults (i.e., infections, traumas, tumoral transformation and radiation) are based on an intricate network of cells and chemical messengers. Abnormally high inflammation immediately after insult or abnormally prolonged pro-inflammatory stimuli bringing about chronic inflammation can lead to life-threatening or severely debilitating diseases. Mesenchymal stem cell (MSC) transplant has proved to be an effective therapy in preclinical studies which evaluated a vast diversity of inflammatory conditions. MSCs lead to resolution of inflammation, preparation for regeneration and actual regeneration, and then ultimate return to normal baseline or homeostasis. However, in clinical trials of transplanted MSCs, the expectations of great medical benefit have not yet been fulfilled. As a practical alternative to MSC transplant, a synthetic drug with the capacity to boost endogenous MSC expansion and/or activation may also be effective. Regarding this, IMT504, the prototype of a major class of immunomodulatory oligonucleotides, induces in vivo expansion of MSCs, resulting in a marked improvement in preclinical models of neuropathic pain, osteoporosis, diabetes and sepsis. IMT504 is easily manufactured and has an excellent preclinical safety record. In the small number of patients studied thus far, IMT504 has been well-tolerated, even at very high dosage. Further clinical investigation is necessary to demonstrate the utility of IMT504 for resolution of inflammation and regeneration in a broad array of human diseases that would likely benefit from an immunoprotective/immunoregenerative therapy.Fil: Zorzopulos, Jorge. Immunotech; ArgentinaFil: Opal, Steven M.. Memorial Hospital of Rhode Island; Estados Unidos. Alpert Medical School; Estados UnidosFil: Hernando Insúa, Andrés. Fundación Pablo Cassara; ArgentinaFil: Rodriguez, Juan M.. Fundación Pablo Cassara; ArgentinaFil: Elías, Fernanda. Fundación Pablo Cassara; ArgentinaFil: Fló, Juan. Immunotech; ArgentinaFil: López, Ricardo A.. Imunotech; ArgentinaFil: Chasseing, Norma Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Lux, Victoria Adela R.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Coronel, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Franco, Raul. Imunotech; ArgentinaFil: Montaner, Alejandro D. Fundación Pablo Cassara; ArgentinaFil: Horn, David L. David Horn Llc; Estados Unido

Garbage in, garbage out: how reliable training data improved a virtual screening approach against SARS-CoV-2 MPro

Introduction: The identification of chemical compounds that interfere with SARS-CoV-2 replication continues to be a priority in several academic and pharmaceutical laboratories. Computational tools and approaches have the power to integrate, process and analyze multiple data in a short time. However, these initiatives may yield unrealistic results if the applied models are not inferred from reliable data and the resulting predictions are not confirmed by experimental evidence.Methods: We undertook a drug discovery campaign against the essential major protease (MPro) from SARS-CoV-2, which relied on an in silico search strategy –performed in a large and diverse chemolibrary– complemented by experimental validation. The computational method comprises a recently reported ligand-based approach developed upon refinement/learning cycles, and structure-based approximations. Search models were applied to both retrospective (in silico) and prospective (experimentally confirmed) screening.Results: The first generation of ligand-based models were fed by data, which to a great extent, had not been published in peer-reviewed articles. The first screening campaign performed with 188 compounds (46 in silico hits and 100 analogues, and 40 unrelated compounds: flavonols and pyrazoles) yielded three hits against MPro (IC50 ≤ 25 μM): two analogues of in silico hits (one glycoside and one benzo-thiazol) and one flavonol. A second generation of ligand-based models was developed based on this negative information and newly published peer-reviewed data for MPro inhibitors. This led to 43 new hit candidates belonging to different chemical families. From 45 compounds (28 in silico hits and 17 related analogues) tested in the second screening campaign, eight inhibited MPro with IC50 = 0.12–20 μM and five of them also impaired the proliferation of SARS-CoV-2 in Vero cells (EC50 7–45 μM).Discussion: Our study provides an example of a virtuous loop between computational and experimental approaches applied to target-focused drug discovery against a major and global pathogen, reaffirming the well-known “garbage in, garbage out” machine learning principle

La definición del arte antes (y después) de su indefinibilidad

En este trabajo se discuten algunas propuestas de la definibilidad del arte (Weltz, Danto, Dickley, Levinson) para concluir que todas implican alguna forma de "instruccionalismo", dan lugar a una definición "procedimental" que carece del interés y la aplicabilidad de las auténticas definiciones. Se propone una estrategia si suponemos que las últimas manifestaciones del arte revelan una dificultad que está ..

Jorge Luis Borges traductor de Die Verwandlung (Fechas, textos, conjeturas)

Este trabalho se propõe analisar a discutível autenticidade da tradução d’A metamorfose de Kafka, publicada em 1933 pela Revista de Occidente (Madri), sem nome de autor, e publicada novamente em Buenos Aires em 1938 pela editora Losada que atribuiu essa tradução a Jorge Luis Borges (e muitíssimas vezes reeditada, até o presente, com a mesma atribuição). Uma vez que não dispomos de nenhum documento absolutamente comprobatório que sustente uma conclusão definitiva, e como é possível incluir entre esses documentos insuficientes as delarações de Borges, busca-se interrogar aqui os mais variados indícios – entre os quais o conceito de tradução sustentado por Borges – e lê-los a partir de uma perspectiva que não ignore os jogos borgianos

EEG signatures of elementary composition: disentangling genuine composition and expectancy processes

We present an adaptation of Bemis & Pylkkänen’s (2011) two-word, composition-list paradigm to study elementary composition in Spanish using electroencephalography. Our main objective is to determine if EEG is sensitive enough to detect a composition-related activity and to evaluate whether the expectancy of participants to compose contributes to this signal. Although we found relevant activity at the expected channels and times, we also found putative composition-related activity before the second word onset. Using a threshold-free cluster permutation analysis combined with linear models we show a task progression effect for the composition task that is not present for the list task. In a second experiment we evaluate two-word composition while avoiding differential expectancy effects across stimuli by incorporating all conditions to a single task. In this case, we failed to find any significant composition-related activity. We suggest that the composition-related activity measured with EEG may be at least in part carried by expectancy processes arising from the block design of the experiment. We conclude that for this paradigm it is important to complement the block based experiments with those controlling for expectancy, and to use powerful data-driven techniques to analyze the data

A high dose of IMT504, the PyNTTTTGT prototype immunostimulatory oligonucleotide, does not alter embryonic development in rats

Synthetic oligodeoxynucleotides (ODNs) are currently being evaluated as vaccine adjuvants for inducing protective immunity. As maternal vaccination is becoming increasingly common, the potential risk of vaccine formulation using ODN adjuvants should be warranted. A recent study performed in mice suggests that exposure to CpG motifs during pregnancy could result (although at very high doses as compared to the ones proposed for human vaccination) in fetal loss and morphological defects. PyNTTTTGT ODNs are immunostimulatory ODNs not bearing CpG motifs, which are very efficient vaccine adjuvants. In this report, we analyzed the potential teratogenic effect of its prototype IMT504 in rats. This animal model was chosen because PyNTTTTGT ODNs are barely active in mice. Intraperitoneal injection of IMT504 at a dose of 20 mg/kg (more than 1000 times higher than the one proposed for a vaccine dose in humans) at day 6 of pregnancy did not produce a significant decrease in the mean number of implanted fetuses or in the number of live pups delivered. Neither the fetuses nor the offspring presented malformations.Fil: Hernando Insúa, Andrés. Immunotech; ArgentinaFil: Rodriguez, Juan M.. Immunotech; ArgentinaFil: Elías, Fernanda. Immunotech; ArgentinaFil: Fló, Juan. Immunotech; ArgentinaFil: López, Ricardo. Immunotech; ArgentinaFil: Franco, Raul. Immunotech; ArgentinaFil: Lago, Nestor. Gema Biotech; ArgentinaFil: Zorzopulos, Jorge. Immunotech; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Montaner, Alejandro Daniel. Immunotech; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencia y Tecnología "Dr. César Milstein". Fundación Pablo Cassará. Instituto de Ciencia y Tecnología ; Argentin

PyNTTTTGT and CpG immunostimulatory oligonucleotides: effect on granulocyte/monocyte colony-stimulating factor (GM-CSF) secretion by human CD56+ (NK and NKT) cells

CD56+ cells have been recognized as being involved in bridging the innate and acquired immune systems. Herein, we assessed the effect of two major classes of immunostimulatory oligonucleotides (ODNs), PyNTTTTGT and CpG, on CD56+ cells. Incubation of human peripheral blood mononuclear cells (hPBMC) with some of these ODNs led to secretion of significant amounts of interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α) and granulocyte/monocyte colony-stimulating factor (GM-CSF), but only if interleukin 2 (IL2) was present. IMT504, the prototype of the PyNTTTTGT ODN class, was the most active. GM-CSF secretion was very efficient when non-CpG ODNs with high T content and PyNTTTTGT motifs lacking CpGs were used. On the other hand, CpG ODNs and IFNα inhibited this GM-CSF secretion. Selective cell type removal from hPBMC indicated that CD56+ cells were responsible for GM-CSF secretion and that plasmacytoid dendritic cells (PDCs) regulate this process. In addition, PyNTTTTGT ODNs inhibited the IFNα secretion induced by CpG ODNs in PDCs by interference with the TLR9 signaling pathway. Since IFNα is essential for CD56+ stimulation by CpG ODNs, there is a reciprocal interference of CpG and PyNTTTTGT ODNs when acting on this cell population. This suggests that these synthetic ODNs mimic different natural alarm signals for activation of the immune system.Fil: Rodriguez, Juan M.. Fundacion Pablo Cassara; ArgentinaFil: Marchicio, José. Immunotech S.a.. Departamento de Investigacion y Desarrollo; ArgentinaFil: López, Mariela. Immunotech S.a.. Departamento de Investigacion y Desarrollo; ArgentinaFil: Ziblat, Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Immunotech S.a.; ArgentinaFil: Elias, Fernanda. Immunotech S.a.. Departamento de Investigacion y Desarrollo; ArgentinaFil: Fló, Juan. Immunotech S.a.. Departamento de Investigacion y Desarrollo; ArgentinaFil: López, Ricardo A.. Immunotech S.a.. Departamento de Investigacion y Desarrollo; ArgentinaFil: Horn, David. David Horn LLC; Estados UnidosFil: Zorzopulos, Jorge. Immunotech S.a.. Departamento de Investigacion y Desarrollo; ArgentinaFil: Montaner, Alejandro Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencias y Tecnología "Dr. Cesar Milstein"; Argentin

IMT504, the prototype of the immunostimulatory oligonucleotides of the PyNTTTTGT class, increases the number of progenitors of mesenchymal stem cells both in vitro and in vivo: potential use in tissue repair therapy

Bone marrow (BM)-derived adult mesenchymal stem cells (MSCs) have the capacity to differentiate in vitro into different cell lines. This makes them a likely source for application in tissue repair therapies. Here, we report evidence indicating that, both in vivo and in vitro, IMT504, the prototype of the PyNTTTTGT class of immunostimulatory oligonucleotides, significantly increases the number of fibroblast colony-forming units (CFU-Fs) that originate MSCs. When rat BM cells were cultured with IMT504, the mean number of CFU-Fs increased about three times as compared with untreated controls (CFU-F: 19 +/- 6.3 vs. 6.8 +/- 2.0/2 x 10(6) seeded BM cells, p = .03). Furthermore, rats inoculated with IMT504 had a significantly higher number of CFU-Fs both in BM (CFU-F: 124 +/- 33 vs. 38 +/- 17/femur, p = .04) and in peripheral blood (animals with detectable CFU-Fs in circulation 8/12 vs. 2/12, p = .04) as compared with untreated animals. On the other hand, BM-derived adherent cells either treated in vitro with IMT504 or obtained from animals injected with IMT504 possess the capacity to differentiate to the osteogenic and adipogenic cell lineages as regular MSCs. Finally, we found that repair of a bone defect was accelerated in rats injected with IMT504 as compared with control animals (area with consolidated bone: 80% +/- 6.4% vs. 49% +/- 3.5%, p = .03, n = 10 rats per group). Importantly, when two human BM were cultured in the presence of IMT504, the mean number of fibroblastic adherent colonies also increased as compared with controls. These results suggest the possibility of clinical use of IMT504 in bone, and presumably other, tissue repair therapies.Fil: Hernando Insúa, Andrés. Immunotech S.a.; ArgentinaFil: Montaner, Alejandro Daniel. Fundación Pablo Cassara; ArgentinaFil: Rodriguez, Juan Manuel. Immunotech S.a.; ArgentinaFil: Elías, Fernanda. Immunotech S.a.; ArgentinaFil: Fló, Juan. Immunotech S.a.; ArgentinaFil: López, Ricardo A.. Immunotech S.a.; ArgentinaFil: Zorzopulos, Ricardo A.. Immunotech S.a.; ArgentinaFil: Hofer, Erica L.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Chasseing, Norma Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Addition of the immunostimulatory oligonucleotide IMT504 to a seasonal flu vaccine increases hemagglutinin antibody titers in young adult and elder rats, and expands the anti-hemagglutinin antibody repertoire

Fil: Montaner, Alejandro Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Centro de Ciencia y Tecnología "Dr. Cesar Milstein"; Argentina.Fil: Denichilo, Analía. Fundación Pablo Cassará; Argentina.Fil: Rodríguez, Juan Manuel. Fundación Pablo Cassará; Argentina.Fil: Fló, Juan. Immunotech S.A.; Argentina.Fil: López, Ricardo Agustin. Immunotech S.A.; Argentina.Fil: Pontoriero, Andrea. ANLIS Dr.C.G.Malbrán. Instituto de Enfermedades Infecciosas; Argentina.Fil: Savy, Vilma. ANLIS Dr.C.G.Malbrán. Instituto de Enfermedades Infecciosas; Argentina.Fil: Baumeister, Elsa. ANLIS Dr.C.G.Malbrán. Instituto de Enfermedades Infecciosas; Argentina.Fil: Frank, Ronald. Helmholtz Centre for Infection Research; Braunschweig, Alemania.Fil: Zorzopulos, Jorge. Fundación Pablo Cassará; Argentina.Fil: Elías, Fernanda. Fundación Pablo Cassará; Argentina.Flu vaccines are partially protective in infants and elder people. New adjuvants such as immunostimulatory oligonucleotides (ODNs) are strong candidates to solve this problem, because a combination with several antigens has demonstrated effectiveness. Here, we report that IMT504, the prototype of a major class of immunostimulatory ODNs, is a potent adjuvant of the influenza vaccine in young adult and elderly rats. Flu vaccines that use virosomes or whole viral particles as antigens were combined with IMT504 and injected in rats. Young adult and elderly animals vaccinated with IMT504-adjuvated preparations reached antibody titers 20-fold and 15-fold higher than controls, respectively. Antibody titers remained high throughout a 120 day-period. Animals injected with the IMT504-adjuvated vaccine showed expansion of the anti-hemagglutinin antibody repertoire and a significant increase in the antibody titer with hemagglutination inhibition capacity when confronted to viral strains included or not in the vaccine. This indicates that the addition of IMT504 in flu vaccines may contribute to the development of significant cross-protective immune response against shifted or drifted flu strains