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Tables show PK kinetics, plasma cell numbers, and patient data. Methods include all supplemental methods.

Trop-2, also known as TACSTD2, EGP-1, GA733-1, and M1S1, is frequently expressed on a variety of human carcinomas, and its expression is often associated with poor prognosis of the diseases. However, it is also present on the epithelium of several normal tissues. A comprehensively designed Trop-2–targeting antibody–drug conjugate (ADC), balancing both efficacy and toxicity, is therefore necessary to achieve clinical utility. To this end, we developed a cleavable Trop-2 ADC (RN927C) using a site-specific transglutaminase-mediated conjugation method and a proprietary microtubule inhibitor (MTI) linker-payload, PF-06380101. Robust in vitro cytotoxicity of RN927C was observed on a panel of Trop-2–expressing tumor cell lines, with IC50 generally in the subnanomolar range. As expected for an MTI-containing ADC, RN927C readily induced mitotic arrest of treated cells in vitro and in vivo, followed by subsequent cell death. The in vivo efficacy of RN927C was tested in multiple cell line and patient-derived xenograft tumor models, including pancreatic, lung, ovarian, and triple-negative breast tumor types. Single-dose administration of RN927C at 0.75 to 3 mg/kg was generally sufficient to induce sustained regression of Trop-2–expressing tumors and showed superior efficacy over standard treatment with paclitaxel or gemcitabine. Administration of RN927C in nonhuman primate toxicity studies resulted in target-mediated effects in skin and oral mucosa, consistent with Trop-2 expression in these epithelial tissues with minimal, non–dose limiting off-target toxicities. On the basis of the combined efficacy and safety results, RN927C is postulated to have a favorable therapeutic index for treatment of solid tumors. Mol Cancer Ther; 15(11); 2698–708. ©2016 AACR.

BCMA bispecific is activity in the presence of soluble BCMA and APRIL.

The restricted expression pattern of B-cell maturation antigen (BCMA) makes it an ideal tumor-associated antigen (TAA) for the treatment of myeloma. BCMA has been targeted by both CD3 bispecific antibody and antibody–drug conjugate (ADC) modalities, but a true comparison of modalities has yet to be performed. Here we utilized a single BCMA antibody to develop and characterize both a CD3 bispecific and 2 ADC formats (cleavable and noncleavable) and compared activity both in vitro and in vivo with the aim of generating an optimal therapeutic. Antibody affinity, but not epitope was influential in drug activity and hence a high-affinity BCMA antibody was selected. Both the bispecific and ADCs were potent in vitro and in vivo, causing dose-dependent cell killing of myeloma cell lines and tumor regression in orthotopic myeloma xenograft models. Primary patient cells were effectively lysed by both CD3 bispecific and ADCs, with the bispecific demonstrating improved potency, maximal cell killing, and consistency across patients. Safety was evaluated in cynomolgus monkey toxicity studies and both modalities were active based on on-target elimination of B lineage cells. Distinct nonclinical toxicity profiles were seen for the bispecific and ADC modalities. When taken together, results from this comparison of BCMA CD3 bispecific and ADC modalities suggest better efficacy and an improved toxicity profile might be achieved with the bispecific modality. This led to the advancement of a bispecific candidate into phase I clinical trials.

Supplementary Figure S1.RN927C induced mitotic arrest of pancreatic PDX tumors in vivo; Supplementary Figure S2. Unconjugated anti-Trop-2 antibody reduced RN927C in vivo efficacy

Pelvic radiation therapy (RT) is a common treatment for malignancies, including gynecological, genitourinary, and lower gastrointestinal tract cancers. However, chronic radiation proctitis (RP) is an unavoidable side effect, and its clinical presentation varies from asymptomatic to potentially life-threatening. This study evaluated the clinical characteristics and risk factors of chronic RP.Patients with chronic RP (212) following RT for various pelvic cancers between January 2015 and December 2021 were enrolled. Clinical characteristics of RP were analyzed retrospectively. Severity was graded according to the Radiation Therapy Oncology Group (RTOG) modified rectal toxicity score and Vienna rectoscopy score (VRS), and risk factors were statistically analyzed.The most common pelvic cancer observed was cervical cancer. The patients received three-dimensional conformal RT (3D-CRT), intensity-modulated RT, or a combination of 3D-CRT and intracavitary RT (ICR). Rectal bleeding occurred in 70 (33.0%) patients. Previous abdominopelvic surgery and total radiation dose significantly correlated with the RTOG score and VRS. Previous abdominopelvic surgery, ICR, and total radiation dose were associated with chronic hemorrhagic RP. All patients with chronic hemorrhagic RP were treated with argon plasma coagulation (APC). 91.4% of cases required 1-3 APC sessions to resolve the bleeding, with a mean of 1.7 sessions.Our results showed that previous abdominopelvic surgery and total radiation dose were significant risk factors related to chronic RP, while total radiation dose was related to chronic hemorrhagic RP. We also showed that APC was effective and safe for chronic hemorrhagic RP.

CD47 is an immunoregulatory protein that is found on the cell surface and plays significant roles in cellular functions such as proliferation, apoptosis, migration, and immune homeostasis. CD47 is overexpressed in various human cancers and is associated with tumor development, progression, and poor prognosis. In this study, we analyzed the expression of CD47 to determine whether it affected the oncogenic behavior of colorectal cancer (CRC) and investigated the prognostic value of CD47 expression in patients with CRC. We investigated 468 patients with CRC who underwent curative surgery and examined the expression of CD47 in tumor and lymph node tissues by performing RT-PCR and immunohistochemistry. Apoptosis, proliferation, angiogenesis, and lymphangiogenesis were determined via a TUNEL assay and immunohistochemical staining for Ki-67, CD34, and D2-40. CD47 expression was increased in human CRC tumors and metastatic lymph nodes compared with normal colorectal mucosa and non-metastatic lymph node tissues. CD47 expression was significantly associated with perineural invasion, lymphovascular invasion, cell differentiation, cancer stage, depth of invasion, lymph node metastasis, distant metastasis, and poor survival. The mean apoptotic index and microvessel density value of CD47-positive tumors were significantly higher than those of CD47-negative tumors. However, no significant difference was observed between CD47 expression and Ki-67 labeling index or lymphatic vessel density. These results indicate that CD47 mediated the progression of CRC by inducing tumor cell apoptosis and angiogenesis.

Background Toll-like receptor (TLR) pathways play a crucial role in mounting potent innate immune responses against invading pathogens, as well as the subsequent engagement of adaptive immunity. Innate immune activation via the TLR9 pathway has potential for treating cancer as demonstrated clinically with TLR9 agonists administered intra-tumorally in melanoma patients.1 We developed a novel toll-like receptor agonist antibody conjugate (TRAAC) platform to systemically deliver a differentiated, targeted TLR9 agonist (T-CpG) for immune activation. The activation of TLR9 pathways can be directed systemically towards specific immune cell populations and tumor microenvironment via antibodies binding to various immune cell receptors. Using multiple TRAACs targeting either immune cells including plasmacytoid DCs (pDCs), myeloid and B lymphocytes, or tumor specific antigens, we evaluated immune modulatory phenotypes, therapeutic potentials, as well as safety and tolerability of this platform in pre-clinical settings. Methods TRAACs were generated using site-specific conjugation. In vitro activity of immune- and tumor-targeted antibody-CpG conjugates was evaluated using human PBMCs. Anti-tumor efficacy and mechanistic assessment of B lymphocyte and myeloid cell-targeted antibody-CpG conjugates were conducted in syngeneic tumor models. Pharmacokinetic (PK), pharmacodynamic and exploratory toxicity evaluations were performed in non-human primates (NHP). Results T-CpG is comprised of monomeric CpG-containing oligonucleotides optimized for potency and stability as an antibody conjugate. TRAACs targeting immune cells enable directed TLR9 activation leading to potent cytokine production and cellular activation that is superior to free CpG. This targeted immune activation also elicits a cascade of downstream modulation of non-targeted immune cells. When administrated systemically in multiple syngeneic models, murine TRAACs targeting either immune cells or tumor antigens exhibited potent, durable, and dose-dependent anti-tumor activity as a single agent and in combination with T-cell checkpoint inhibitors (CPIs). A single peripheral dose of either B lymphocyte or myeloid targeted-CpG evoked both innate and adaptive immune responses within the tumor microenvironment as demonstrated by NanoString analysis. The observed immunomodulatory phenotypes are consistent with those elicited by direct intra-tumoral CpG delivery. Following repeated intravenous doses in NHP, TRAACs demonstrated targeted receptor occupancy, antibody-like PK, and favorable tolerability profile. Conclusions Pre-clinical evaluation of a novel platform comprised of antibodies conjugated to a differentiated TLR9 agonist demonstrated targeted immune activation, potent anti-tumor activity as single agent and in combination with CPIs and favorable tolerability profiles in NHPs. Such antibody-CpG conjugates have the potential for clinical development as systemically delivered therapeutics providing powerful innate and adaptive anti-tumor immunity across multiple tumor types. Reference Hamid O, Ismail R, Puzanov I. Intratumoral Immunotherapy—Update 2019. The Oncol2019; 25:343–359.

Rationale: Syphilis is a contagious infectious disease caused by Treponema pallidum. Gastric involvement of syphilis is rare and has nonspecific gastrointestinal symptoms and endoscopic findings. To date, 16 cases have been reported in Korea. Here, we report 2 additional cases of gastric syphilis in men in their 30 second. Patients concerns: Two 35- and 33-year-old men presented with epigastric pain. Diagnosis: The serum venereal disease research laboratory and fluorescent treponemal antibody absorption tests were positive. Esophagogastroduodenoscopy showed multiple variable-sized flat elevated lesions and geographic ulcers with whitish exudates in the antrum and body. Warthin–Starry silver staining of endoscopic biopsy specimens confirmed gastric syphilis. Interventions: The patients were treated with an intramuscular injection of 2.4 million units of benzathine penicillin once a week for 3 weeks. Outcomes: Clinical symptoms and gastric lesions were completely resolved. Lessons: First, gastric syphilis, despite its rarity and nonspecific symptoms and endoscopic findings, should be considered in a rare extracutaneous presentation of syphilis. Second, a high index of clinical suspicion and an accurate diagnosis based on a combination of clinical, radiological, endoscopic, serologic, and histopathologic findings provide an opportunity to identify and treat patients with gastric syphilis.

There has been increased use of self-expandable metal stents (SEMS) in treating malignant colorectal obstruction (MCO). The aim of this study was to investigate factors that are associated with the outcomes of SEMS placement for MCO.Clinical data from patients who underwent SEMS placement for MCO at 6 hospitals in Honam province of South Korea between 2009 and 2018 were reviewed retrospectively. Eight hundred two patients were identified and their data were analyzed. Technical success, clinical success, complications, and predictors of outcome were included as main outcome measures.Technical and clinical success rates were 98.8% (792/802) and 90.1% (723/802), respectively. Complications including stent migration, stent occlusion due to tumor ingrowth and outgrowth, perforation, bacteremia/fever, and bleeding occurred in 123 (15.3%) patients. In multivariate regression analyses, procedure time was significantly associated with the technical success of SEMS placement (P = .001). Longer length of obstruction, the use of covered stent, and longer procedure time were significant independent predictive factors for the clinical success of SEMS placement (odds ratio [OR] 0.974 (95% confidence interval [CI] 0.950-0.990); P = .043, OR 0.255 (95% CI 0.138-0.471); P

Novel therapies capable of engaging both innate and adaptive immune responses may engender durable anti-tumor immunity. Activation of toll-like receptor 9 (TLR9) by unmethylated oligonucleotides promotes innate inflammatory responses and the induction of adaptive immunity. Recently, several CpG-based TLR9 agonists have demonstrated clinical benefit in melanoma patients when administered intra-tumorally. Immune cells expressing TLR9 include B cells, plasmacytoid dendritic cells and myeloid cells. In B cells, TLR9 activation in human and mouse drives B cell proliferation and differentiation. Recent studies have described the presence and potential role of B cells in the response to checkpoint inhibitors in multiple solid tumor types. Specifically, enrichment of memory B cells, plasma cells and tertiary lymphoid organs have been observed in the tumors of responders to checkpoint inhibitors. We developed a novel Toll-like Receptor Agonist Antibody Conjugate (TRAAC) platform to deliver a potent and differentiated TLR9 agonist (T-CpG) for targeted immune activation via systemic administration. TAC-001 is comprised of T-CpG conjugated to an anti-CD22 antibody, a receptor restricted to B cells. It is designed to systemically deliver T-CpG to B cells by binding CD22 receptor on B cells, leading to TLR9 signaling, B cell activation and a cascade of immune reactions. We show that the innate and adaptive immune responses triggered by TAC-001 leads to potent anti-tumor activity. In vitro stimulation of B cells with TAC-001 leads to increase expression of co-stimulatory molecules, cross-presentation leading to T cell proliferation and induction of immunoglobulin secretion. In mice, systemic delivery of a murine reactive anti-CD22-CpG conjugate shows durable single agent activity in PD-1 refractory and PD-1 resistant tumors. Mice with eradicated tumors acquire memory immune response upon tumor rechallenge. The cellular response in mice treated with TAC-001 exhibit B cell differentiation with significant decrease in IL-10+Bregs, along with increased in T cell effector function and modulation in suppressive myeloid cells within the tumor microenvironment. Systemic intravenous administration of TAC-001 in monkeys showed favorable tolerability, PK and PD profiles. These results demonstrate the unique properties of TAC-001 which integrates TLR9 activation and B cells to employ both innate and adaptive immune responses to promote anti-tumor activity. These data support the development of TAC-001 for a broad range of solid tumor malignancies. Citation Format: Tracy C. Kuo, Ons Harrabi, Amy Chen, Emma R. Sangalang, Laura Doyle, Danielle Fontaine, Min Li, Bora Han, Jaume Pons, Janet Sim, Hong I. Wan. TAC-001, a toll-like receptor 9 (TLR9) agonist antibody conjugate targeting B cells, promotes anti-tumor immunity and favorable safety profile following systemic administration in preclinical models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1721.

The restricted expression pattern of B-cell maturation antigen (BCMA) makes it an ideal tumor-associated antigen (TAA) for the treatment of myeloma. BCMA has been targeted by both CD3 bispecific antibody and antibody–drug conjugate (ADC) modalities, but a true comparison of modalities has yet to be performed. Here we utilized a single BCMA antibody to develop and characterize both a CD3 bispecific and 2 ADC formats (cleavable and noncleavable) and compared activity both in vitro and in vivo with the aim of generating an optimal therapeutic. Antibody affinity, but not epitope was influential in drug activity and hence a high-affinity BCMA antibody was selected. Both the bispecific and ADCs were potent in vitro and in vivo, causing dose-dependent cell killing of myeloma cell lines and tumor regression in orthotopic myeloma xenograft models. Primary patient cells were effectively lysed by both CD3 bispecific and ADCs, with the bispecific demonstrating improved potency, maximal cell killing, and consistency across patients. Safety was evaluated in cynomolgus monkey toxicity studies and both modalities were active based on on-target elimination of B lineage cells. Distinct nonclinical toxicity profiles were seen for the bispecific and ADC modalities. When taken together, results from this comparison of BCMA CD3 bispecific and ADC modalities suggest better efficacy and an improved toxicity profile might be achieved with the bispecific modality. This led to the advancement of a bispecific candidate into phase I clinical trials.

Trop-2, also known as TACSTD2, EGP-1, GA733-1, and M1S1, is frequently expressed on a variety of human carcinomas, and its expression is often associated with poor prognosis of the diseases. However, it is also present on the epithelium of several normal tissues. A comprehensively designed Trop-2–targeting antibody–drug conjugate (ADC), balancing both efficacy and toxicity, is therefore necessary to achieve clinical utility. To this end, we developed a cleavable Trop-2 ADC (RN927C) using a site-specific transglutaminase-mediated conjugation method and a proprietary microtubule inhibitor (MTI) linker-payload, PF-06380101. Robust in vitro cytotoxicity of RN927C was observed on a panel of Trop-2–expressing tumor cell lines, with IC50 generally in the subnanomolar range. As expected for an MTI-containing ADC, RN927C readily induced mitotic arrest of treated cells in vitro and in vivo, followed by subsequent cell death. The in vivo efficacy of RN927C was tested in multiple cell line and patient-derived xenograft tumor models, including pancreatic, lung, ovarian, and triple-negative breast tumor types. Single-dose administration of RN927C at 0.75 to 3 mg/kg was generally sufficient to induce sustained regression of Trop-2–expressing tumors and showed superior efficacy over standard treatment with paclitaxel or gemcitabine. Administration of RN927C in nonhuman primate toxicity studies resulted in target-mediated effects in skin and oral mucosa, consistent with Trop-2 expression in these epithelial tissues with minimal, non–dose limiting off-target toxicities. On the basis of the combined efficacy and safety results, RN927C is postulated to have a favorable therapeutic index for treatment of solid tumors. Mol Cancer Ther; 15(11); 2698–708. ©2016 AACR.

CD47 is a widely expressed cell surface protein that functions as an immune checkpoint in cancer. When expressed by tumor cells, CD47 can bind SIRPα on myeloid cells, leading to suppression of tumor cell phagocytosis and other innate immune functions. CD47-SIRPα signaling has also been implicated in the suppression of adaptive antitumor responses, but the relevant cellular functions have yet to be elucidated. Therapeutic blockade of the CD47 pathway may stimulate antitumor immunity and improve cancer therapy. To this end, a novel CD47-blocking molecule, ALX148, was generated by fusing a modified SIRPα D1 domain to an inactive human IgG1 Fc. ALX148 binds CD47 from multiple species with high affinity, inhibits wild type SIRPα binding, and enhances phagocytosis of tumor cells by macrophages. ALX148 has no effect on normal human blood cells in vitro or on blood cell parameters in rodent and non-human primate studies. Across several murine tumor xenograft models, ALX148 enhanced the antitumor activity of different targeted antitumor antibodies. Additionally, ALX148 enhanced the antitumor activity of multiple immunotherapeutic antibodies in syngeneic tumor models. These studies revealed that CD47 blockade with ALX148 induces multiple responses that bridge innate and adaptive immunity. ALX148 stimulates antitumor properties of innate immune cells by promoting dendritic cell activation, macrophage phagocytosis, and a shift of tumor-associated macrophages toward an inflammatory phenotype. ALX148 also stimulated the antitumor properties of adaptive immune cells, causing increased T cell effector function, pro-inflammatory cytokine production, and a reduction in the number of suppressive cells within the tumor microenvironment. Taken together, these results show that ALX148 binds and blocks CD47 with high affinity, induces a broad antitumor immune response, and has a favorable safety profile.

L’etude des particularites visuelles dans les Troubles du spectre autistique (TSA) pose une question centrale, celle du caractere general de ses particularites ou de leur limitation au traitement des stimuli socio-emotionnels. Une comparaison systematique entre le traitement de differents types de stimuli sous-tend la reponse a cette question. Cependant jusqu’a present les comparaisons realisees ont porte sur des stimuli statiques, et lorsque des stimuli dynamiques ont ete utilises, ils n’ont pas porte de facon comparable sur les mouvements biologiques du visage (expressions emotionnelles) et sur les mouvements physiques des objets. Ayant mis au point un dispositif permettant de comparer systematiquement le changement d’etat des visages emotionnels, des robots emotionnels et des objets, (http://www.lutin-userlab.fr/FOT/), nous avons conduit une experimentation combinant la technique du morphing et de l’eye-Tracking. Douze adolescents non-verbaux ont ete apparies a deux groupes typiques : l’un forme d’enfants de meme âge developpemental, l’autre d’adolescents de meme âge chronologique. Les resultats montrent une asymetrie de traitement perceptif selon les stimuli : intact pour les stimuli physiques, le traitement apparait deficitaire pour les stimuli emotionnels dans le cas de participants de bas niveau cognitif avec TSA. Ces resultats sont en faveur d’un dysfonctionnement de traitement perceptif concernant de facon specifique le domaine socio-emotionnel.

Targeting the CD47-SIRPα myeloid checkpoint pathway represents a novel therapeutic approach to enhance anti-cancer immunity. CD47 is a widely expressed cell surface protein that functions as a marker of self. Signal regulatory protein-α (SIRPα) is an ITIM-containing inhibitory receptor of CD47 on myeloid cells. Tumor cells up-regulate CD47 expression to evade immune surveillance. Clinical trials targeting CD47 to inhibit SIRPα interaction have shown promising results (N Lakhani et al, 2018 SITC #P335). Unlike CD47, which is expressed ubiquitously, SIRPα has limited expression, mainly on myeloid cells including macrophages, dendritic cells, and neutrophils, and neurons. Targeting SIRPα circumvents the need to overcome the CD47 sink that is necessary for CD47 targeting molecules. Antibodies to SIRPα that bind mouse, human and cynomolgus monkey SIRPα with high affinity, were evaluated for their effects on macrophage function, anti-tumor activity and normal blood cells. Humanized antibody to SIRPα was generated to bind both human SIRPα allele V1 and V2 with high affinity (pM) and cross-react with cynomolgus and mouse SIRPα. Anti-SIRPα antibody in combination with targeted anti-cancer antibodies enhanced antibody-dependent cellular phagocytosis (ADCP) of tumor cell lines by human monocyte derived macrophages. In preclinical xenograft models of SIRPα positive and negative malignant cell lines, administration of anti-SIRPα antibody enhanced anti-tumor activity of rituximab and cetuximab via ADCP. The impact of SIRPα blockade on anti-tumor immunity was assessed in immunocompetent tumor mouse models. SIRPα antibodies enhanced anti-tumor activity of checkpoint inhibitors in syngeneic tumor models. Combination therapy with SIRPα antibodies and checkpoint inhibitors in tumor models showed eradication of tumors, acquired memory immune response upon tumor rechallenge and decreased lung metastasis. The cellular mechanisms driving efficacy include dendritic cell activation, reduction of myeloid suppression and enhancement of T cell effector profile. Exploratory safety and PK/PD profiles of anti-SIRPα antibodies were assessed in cynomolgus monkeys. Antibodies to SIRPα demonstrated a typical antibody PK and complete SIRPα target occupancy. There were no adverse clinical observations, no changes in hematological parameters and no impact on serum chemistries. In summary, anti-SIRPα antibodies combined with targeted anti-cancer antibodies or checkpoint inhibitors integrate innate and adaptive immune responses to promote anti-tumor activity in preclinical models. Together with the favorable safety profile in cynomolgus monkeys, these data support the development of SIRPα antibodies for solid tumors and hematological malignancies. Citation Format: Tracy C. Kuo, Amy Chen, Ons Harrabi, Steven E. Kauder, Emma R. Sangalang, Laura Doyle, Sangeetha Bollini, Bora Han, Janet Sim, Jaume Pons, Hong I. Wan. Antibodies to Sirpα enhance innate and adaptive immune responses to promote anti-tumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 562.

Dexamethasone was given in 2 oral dosing regimens with repeat dose oral administration of the gamma secretase inhibitor (GSI), PF-03084014, in Sprague-Dawley (SD) rats in order to evaluate the effects of coadministration of dexamethasone on GSI-induced goblet cell hyperplasia (GCH) in the intestinal tract. Safety end points were evaluated in 1 week and 1 month studies. The dosing regimens tested in the 1-month studies included a 1-week pretreatment with 1.0 mg/kg dexamethasone followed by a 3-week repeat dose treatment with 100 mg/kg GSI or concurrent intermittent treatment with 1.0 mg/kg dexamethasone on weeks 1 and 3 and repeat dose treatment with 100 mg/kg GSI for 4 weeks. Pretreatment with dexamethasone for 1 week transiently mitigated the severity of intestinal GCH for up to 1 week. Intermittent coadministration of dexamethasone on weeks 1 and 3 with GSI repeat dosing for 4 weeks mitigated intestinal GCH for up to 4 weeks post treatment. Treatment-related morbidity and mortality occurred on day 7 with 150 mg/kg GSI and 5 mg/kg dexamethasone coadministration, and on days 13, 14, and 23 with 100 mg/kg GSI and 1 mg/kg dexamethasone coadministration.

EnglishA one-year pilot study was conducted with 8 children with autism aged 30 to 84 months. The children were proposed 25 individual sessions of reciprocal imitation throughout the school year. The content of the sessions was adapted to the imitation level of each child, as assessed by our imitation scale including spontaneous imitation and recognition of being imitated in addition to the classical evaluation of instructed imitation. Whatever the initial level, the aim of the program was to gently lead children to alternate the roles of imitator and model, so as to achieve an efficient non verbal communicative means, and to improve their imitation capacities. The Before-After comparison of scores shows a significant increase of PEP-R imitation scores after the training, altogether with a significant increase of positive behaviour at the after evaluation and at home. A significant correlation arises between the increase of communicative imitation and the increase of positive behaviour. The developmental patterns used "a la carte" depending of each child's initial level are reported, some of them in full details so as to describe as closely as possible the work elaborated step by step during the training. Results are presented child by child, given the heterogeneity of initial levels. They show for 6 of the 8 children an important increase of positive behaviour and a significant correlation between the increase of communicative imitation and the increase of positive behaviour. The increase of positive behaviour is also quoted by the parents, thus stressing a trend toward the generalization of the improvement. francaisUne etude pilote a ete menee durant un an avec 8 enfants de 30 a 84 mois ayant recu le diagnostic d'autisme. Les enfants ont beneficie de 25 seances individuelles d'imitation reciproque durant l'annee scolaire. Le contenu des seances etait adapte au niveau d'imitation de chaque enfant, prealablement evalue a l'aide d'un instrument ajoutant l'imitation spontanee et la reconnaissance d'etre imite a la classique evaluation de l'imitation induite. Quel que soit le niveau, l'objectif etait de conduire doucement l'enfant a alterner les roles d'imitateur et de modele, de facon a maitriser un moyen efficace de communication non verbale, tout en enrichissant le repertoire d'actions. La comparaison des scores avant-apres montre une augmentation significative de l'imitation au PEP-R et une augmentation des comportements positifs lors de l'evaluation finale et en famille. Une correlation significative se degage lors de l'evaluation apres entre les progres en imitation communicative et l'augmentation des comportements positifs. Les patrons developpementaux sont presentes enfant par enfant, etant donne l'heterogeneite des niveaux de depart, et certains sont abondamment detailles pour decrire le principe du travail elabore pas a pas durant les entrainements.

Genetic variation in the IL-7 receptor-α ( IL-7R ) gene is associated with susceptibility to human type 1 diabetes (T1D). Here we investigate the therapeutic efficacy and mechanism of IL-7Rα antibody in a mouse model of T1D. IL-7Rα antibody induces durable, complete remission in newly onset diabetic mice after only two to three injections. IL-7 increases, whereas IL-7Rα antibody therapy reduces, the IFN-γ–producing CD4 + (T H 1) and IFN-γ–producing CD8 + T cells. Conversely, IL-7 decreases and IL-7Rα antibody enhances the inhibitory receptor Programmed Death 1 (PD-1) expression in the effector T cells. Programmed Death 1 blockade reversed the immune tolerance mediated by the IL-7Rα antibody therapy. Furthermore, IL-7Rα antibody therapy increases the frequency of regulatory T cells without affecting their suppressor activity. The durable efficacy and the multipronged tolerogenic mechanisms of IL-7Rα antibody therapy suggest a unique disease-modifying approach to T1D.

Resume L’article aborde, dans une perspective developpementale, l’analyse de la perception visuelle des changements d’etat dynamique d’agents animes (visages emotionnels) et d’objets inanimes (objets fabriques et naturel). Il explore l’hypothese de mecanismes distincts de traitement de l’information, refletant des modifications de proprietes intentionnelles ou de structures causales. En particulier, la dynamique des visages nous est familiere, tandis que les objets sont generalement statiques et necessitent une intervention externe pour se mettre en mouvement. De surcroit, on peut distinguer deux types d’objets en mouvement : ceux qui sont lies aux evenements physiques et ceux qui sont lies a nos actions. Dans ce dernier cas, la dynamique nous est plus familiere, et peut s’appuyer sur une simulation se rapportant a une intention d’action. Elle est de ce fait plus proche de la resonance motrice, qui est en jeu lorsque nous reagissons a la mimique d’autrui en « resonant » avec lui. Le traitement dynamique des visages et des objets a usage manuel devrait etre facilite lorsque la simulation est possible, ce qui fait intervenir l’experience. Nous postulons donc un effet de l’âge sur les performances concernant ces deux types de stimuli, mais pas concernant les objets naturels. Pour tester cette hypothese, 72 participants repartis en trois groupes, adultes et enfants de 7-9 et 10-12 ans, se sont vus presenter trois types de stimuli dynamiques obtenus a partir de la technique du morphing. Les participants devaient discriminer l’etat intermediaire du changement et retrouver parmi trois images tests, l’image correspondant a cet etat. Les resultats mettent en evidence un effet significatif de l’âge, de la vitesse et de l’ordre de transformation sur la perception du changement d’etat de visage emotionnel et d’objets.

Site-specific conjugation improves therapeutic index of antibody drug conjugates with high drug loading

Bococizumab is a humanized monoclonal IgG2Δa antibody against proprotein convertase subtilisin/kexin type 9 (PCSK9) for the treatment of hyperlipidemia. The evaluation of potential effects on embryo-fetal development was conducted in the rat. In a pharmacokinetic/pharmacodynamic study bococizumab was administered intravenously to pregnant Sprague–Dawley (SD) rats (n = 8/group) at 0, 10, 30, and 100 mg/kg during organogenesis. Maternal and fetal bococizumab, total cholesterol and HDL concentrations were determined. Bococizumab was well tolerated and there were no effects on ovarian or uterine parameters. Maternal and fetal bococizumab exposure increased with increasing dose, with a corresponding dose-dependent decrease in fetal cholesterol levels. Maternal cholesterol levels were decreased significantly, with reductions that were of a similar magnitude regardless of dose. In the definitive embryo-fetal development study bococizumab was administered to pregnant SD rats (n = 20/group) at 0, 10, 30, and 100 mg/kg and no adverse maternal or developmental effects were observed up to 100 mg/kg. These studies have provided an appropriate and relevant safety assessment of bococizumab in pregnant rats to inform human risk assessment, demonstrating no adverse effects on embryo-fetal development at magnitudes greater than anticipated clinical exposure and in the presence of maximal reductions in maternal cholesterol and dose-dependent reductions in fetal cholesterol.

A morphing procedure has been designed to compare directly the perception of emotional expressions and of moving objects. Morphing tasks were presented to 12 low-functioning teenagers with Autism Spectrum Disorder (LF ASD) compared to 12 developmental age-matched typical children and a group presenting ceiling performance. In a first study, when presented with morphed stimuli of objects and emotional faces, LF ASD showed an intact perception of object change of state together with an impaired perception of emotional facial change of state. In a second study, an eye-tracker recorded visual exploration of morphed emotional stimuli displayed by a human face and a robotic set-up. Facing the morphed robotic stimuli, LF ASD displayed equal duration of fixations toward emotional regions and toward mechanical sources of motion, while the typical groups tracked the emotional regions only. Altogether the findings of the two studies suggest that individuals with ASD process motion rather than emotional signals when facing facial expressions.

Vascular inflammation, infusion reactions, glomerulopathies, and other potentially adverse effects may be observed in laboratory animals, including monkeys, on toxicity studies of therapeutic monoclonal antibodies and recombinant human protein drugs. Histopathologic and immunohistochemical (IHC) evaluation suggests these effects may be mediated by deposition of immune complexes (ICs) containing the drug, endogenous immunoglobulin, and/or complement components in the affected tissues. ICs may be observed in glomerulus, blood vessels, synovium, lung, liver, skin, eye, choroid plexus, or other tissues or bound to neutrophils, monocytes/macrophages, or platelets. IC deposition may activate complement, kinin, and/or coagulation/fibrinolytic pathways and result in a systemic proinflammatory response. IC clearance is biphasic in humans and monkeys (first from plasma to liver and/or spleen, second from liver or spleen). IC deposition/clearance is affected by IC composition, immunomodulation, and/or complement activation. Case studies are presented from toxicity study monkeys or rats and indicate IHC-IC deposition patterns similar to those predicted by experimental studies of IC-mediated reactions to heterologous protein administration to monkeys and other species. The IHC-staining patterns are consistent with findings associated with generalized and localized IC-associated pathology in humans. However, manifestations of immunogenicity in preclinical species are generally not considered predictive to humans.

Two 6-month repeat-dose toxicity studies in cynomolgus monkeys illustrated immune complex–mediated adverse findings in individual monkeys and identified parameters that potentially signal the onset of immune complex–mediated reactions following administration of RN6G, a monoclonal antibody (mAb). In the first study, 3 monkeys exhibited nondose-dependent severe clinical signs accompanied by decreased erythrocytes with increased reticulocytes, neutrophilia, monocytosis, thrombocytopenia, coagulopathy, decreased albumin, azotemia, and increased serum levels of activated complement products, prompting unscheduled euthanasia. Histologically, immunohistochemical localization of RN6G was associated with monkey immunoglobulin and complement components in glomeruli and other tissues, attributable to immune complex disease (ICD). All 3 animals also had anti-RN6G antibodies and decreased plasma levels of RN6G. Subsequently, an investigational study was designed and conducted with regulatory agency input to detect early onset of ICD and assess reversibility to support further clinical development. Dosing of individual animals ceased when biomarkers of ICD indicated adverse findings. Of the 12 monkeys, 1 developed anti-RN6G antibodies and decreased RN6G exposure that preceded elevations in complement products, interleukin-6, and coagulation parameters and decreases in albumin and fibrinogen. All findings in this monkey, except for antidrug antibody (ADA), reversed after cessation of dosing without progressing to adverse sequelae typically associated with ICD.

Ischemic/hypoxic brain damage induced in 7-day-old rats was significantly attenuated in a dose-dependent manner by intracerebral injection of glial cell line-derived neurotrophic factor (GDNF; 2 or 4 microg) within 30 min after the insult. Whereas the great majority of the vehicle-treated animals showed massive infarction involving more than 75% of the affected cerebral hemisphere, GDNF injection resulted in a remarkable reduction in both the incidence and severity of the brain damage (incidence ranging from 76% to 93% in controls to 34% to 64% in the 2.0-microg group and 7% to 29% in 4.0-microg group). The induction of immunoreactive 70-kDa heat shock protein (HSP70) in cerebral cortical neurons was also significantly reduced in GDNF-treated animals as compared to controls. The mechanisms responsible for the neuroprotective effects of GDNF remain unknown, although it has been speculated that these may be endogeneous. The higher expression of GDNF and its mRNA in developing brains may be one of the factors responsible for the relative resistance to ischemia of fetal and neonatal as opposed to adult brains. GDNF may possibly act by protecting against oxidative stress or by scavenging free radicals generated during ischemia. The results of our study strongly suggest that GDNF may prove to be an effective and potent protective agent against perinatal ischemic/hypoxic encephalopathy.

Russian knapweed is a perennial weed found in many parts of the world, including southern California. Chronic ingestion of this plant by horses has been reported to cause equine nigropallidal encephalomalacia (ENE), which is associated with a movement disorder simulating Parkinson's disease (PD). Repin, a principal ingredient purified from Russian knapweed, is a sesquiterpene lactone containing an alpha-methylenebutyrolactone moiety and epoxides and is a highly reactive electrophile that can readily undergo conjugation with various biological nucleophiles, such as proteins, DNA, and glutathione (GSH). We show in this study that repin is highly toxic to C57BL/6J mice and Sprague-Dawley rats and acutely induces uncoordinated locomotion associated with postural tremors, hypothermia, and inability to respond to sonic and tactile stimuli. We also show that repin intoxication reduces striatal and hippocampal GSH and increases total striatal dopamine (DA) levels in mice. Striatal microdialysis in rats, however, has demonstrated a significant reduction of extracellular DA levels. These findings, coupled with the absence of any demonstrable change in striatal DOPAC levels, suggest that repin acts by inhibiting DA release, a hypothesis that is further supported by our demonstration that, in cultured PC12 cells, repin inhibits the release of DA without affecting its uptake. We believe, therefore, that inhibition of DA release represents one of the earliest pathogenetic events in ENE, leading eventually to striatal extracellular DA denervation, oxidative stress, and degeneration of nigrostriatal pathways. Since the neurotoxic effects of repin appear to be mediated via oxidative stress, and since repin is a natural product isolated from a plant in our environment that can cause a movement disorder associated with degeneration of nigrostriatal pathways, clarification of the mechanism of repin neurotoxicity may provide new insights into our understanding of the pathogenesis of PD.

Item does not contain fulltext The interleukin-7 receptor alpha chain (IL-7Ralpha) gene was identified as a top non-major histocompatibility complex-linked risk locus for multiple sclerosis (MS). Recently, we showed that a T helper 1 (T(H)1)-driven, but not a T(H)17-driven, form of MS exhibited a good clinical response to interferon-beta (IFN-beta) therapy. We now demonstrate that high serum levels of IL-7, particularly when paired with low levels of IL-17F, predict responsiveness to IFN-beta and hence a T(H)1-driven subtype of MS. We also show that although IL-7 signaling is neither necessary nor sufficient for the induction or expansion of T(H)17 cells, IL-7 can greatly enhance both human and mouse T(H)1 cell differentiation. IL-7 alone is sufficient to induce human T(H)1 differentiation in the absence of IL-12 or other cytokines. Furthermore, targeting IL-7/IL-7Ralpha is beneficial in experimental autoimmune encephalomyelitis (EAE), a mouse model of MS. Mice treated with IL-7Ralpha-blocking antibodies before or after onset of paralysis exhibited reduced clinical signs of EAE, with reduction in peripheral naive and activated T cells, whereas central memory T, regulatory T, B, and natural killer cell populations were largely spared. IL-7Ralpha antibody treatment markedly reduced lymphocyte infiltration into the central nervous system in mice with EAE. Thus, a serum profile of high IL-7 may signify a T(H)1-driven form of MS and may predict outcome in MS patients undergoing IFN-beta therapy. Blockade of IL-7 and the IL-7Ralpha pathway may have therapeutic potential in MS and other autoimmune diseases.

The vanilloid receptor TRPV1 (transient receptor potential vanilloid 1) is a cation channel that serves as a polymodal detector of pain-producing stimuli such as capsaicin, protons (pH in vivoand that TRPV1 antagonism and hyperthermia are not separable. TRPV1 antagonists caused hyperthermia in multiple species (rats, dogs, and monkeys), demonstrating that TRPV1 function in thermoregulation is conserved from rodents to primates. Together, these results indicate that tonic TRPV1 activation regulates body temperature.

Iron chelators inhibit endotoxin-induced NF-kappaB activation in hepatic macrophages (HMs), suggesting a role for the intracellular chelatable pool of iron in NF-kappaB activation. The present study tested this hypothesis. Analysis of Fe(59)-loaded HMs stimulated with lipopolysaccharide (LPS), revealed a previously unreported, transient rise in intracellular low molecular weight (LMW).Fe(59) complex ([LMW.Fe](i)) at/=2 min returning to the basal level within 15 min. The [LMW.Fe](i) response preceded IkappaB kinase (IKK) (/=15 min) and NF-kappaB (/=30 min) activation. Iron chelators (1,2-dimethyl-3-hydroxypyridin-4-one and N,N'-bis-2-hydroxybenzylethylenediamine-N,N'-diacetic acid) abrogated the [LMW.Fe](i) response and IKK and NF-kappaB activation. The [LMW.Fe](i) response was also observed in tumor necrosis factor alpha (TNFalpha)-stimulated HMs and RAW264.7 cells treated with LPS and interferon-gamma but not in primary rat hepatocytes or myofibroblastic cells exposed to LPS or TNFalpha. Both [LMW.Fe](i) response and IKK activation in LPS-stimulated HMs were inhibited by diphenylene iodonium (nonspecific inhibitor for flavin-containing oxidases), l-N(6)-(1-iminoethyl)lysine (selective iNOS inhibitor), and adenoviral-mediated expression of a dominant negative mutant of Rac1 or Cu,Zn-superoxide dismutase, suggesting the role of (.)NO and O(2)() in mediating the iron signaling. In fact, this inhibition was recapitulated by a cell-permeable scavenger of ONOO(-), 5,10,15,20-tetrakis (4-sulfonatophenyl)porphyrinato iron (III) chloride. Conversely, ONOO(-) alone induced both [LMW.Fe](i) response and IKK activation. Finally, direct addition of ferrous iron to cultured HMs activated IKK and NF-kappaB. These results support a novel signaling role for [LMW.Fe](i) in IKK activation, which appears to be induced by ONOO(-) and selectively operative in macrophages.

Notch signaling is deregulated in T-cell acute lymphoblastic leukemia (T-ALL) and advanced solid tumors, making it an attractive target for oncology drug development. In this present report, we describe a novel mouse monoclonal antibody, Notch1 mAb, that specifically binds to the negative regulatory region (NRR) of human Notch1 receptor. Using a T-ALL cell line, HPB-ALL, that harbors mutations in Notch1 heterodimerization and PEST domains, we demonstrated that Notch1 mAb blocked Notch signaling by reduction of Notch1 intracellular domain (NICD) and down-regulation of Notch target genes, Hes-1 and cMyc. Notch1 mAb caused cell growth inhibition of HPB-ALL and several other T-ALL cell lines, via induction of cell cycle arrest and apoptosis. Notch1 mAb treatment resulted in robust NICD reduction and marked antitumor efficacy in HBP-ALL xenograft model. Additional mechanism-of-action studies revealed inhibition of tumor cell proliferation and induction of apoptosis in HPB-ALL tumors, suggesting that the anti-tumor activity of Notch1 mAb may be mediated by its direct effects on tumor cell growth or survival. Furthermore, this antibody led to a significant reduction in leukemia progenitor cells (LPCs) baring NOTCH1 mutation in bioluminescent humanized T-ALL LPC mouse models. In addition to its inhibitory effect on mutant Notch1, Notch1 mAb also displayed robust neutralization activity on wild type Notch1 and caused tumor growth inhibition in breast cancer models MDA-MB-231 and MX-1 by targeting the wild type receptor in these tumor types. Interestingly, using a gamma secretase inhibitor PF-03084014, we showed that Notch1 mAb and PF-03084014 elicited similar degree of biology responses in HPB-ALL cells. Our results indicate that antibodies that bind to NRR can act as potent inhibitors of Notch1 signaling and provide opportunities for development of novel cancer therapeutics for T-ALL and solid tumors. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1765. doi:10.1158/1538-7445.AM2011-1765