Ruxolitinib, a JAK1/2 Inhibitor, Ameliorates Cytokine Storm in Experimental Models of Hyperinflammation Syndrome.

Hyperinflammatory syndromes comprise a heterogeneous group of disorders characterized by severe inflammation, multiple organ dysfunction, and potentially death. In response to antigenic stimulus (e.g., SARS-CoV-2 infection), overactivated CD8+ T-cells and macrophages produce high levels of proinflammatory cytokines, such as IFN-γ, TNF-α, IL-6, and IL-12. Multiple inflammatory mediators implicated in hyperinflammatory syndromes utilize the Janus kinase-signal transducers and activators of transcription (JAK-STAT) cascade to propagate their biological function. Our findings demonstrate that oral ruxolitinib dosing designed to mimic clinically relevant JAK-STAT pathway inhibition significantly reduces the harmful consequences of immune overactivation in multiple hyperinflammatory models. In contrast to monoclonal antibody therapies targeting a single cytokine, ruxolitinib effectively downregulates the functional effect of multiple cytokines implicated in hyperinflammatory states, without broad immunosuppression.

JAK/STAT pathway inhibition sensitizes CD8 T cells to dexamethasone-induced apoptosis in hyperinflammation.

Cytokine storm syndromes (CSS) are severe hyperinflammatory conditions characterized by excessive immune system activation leading to organ damage and death. Hemophagocytic lymphohistiocytosis (HLH), a disease often associated with inherited defects in cell-mediated cytotoxicity, serves as a prototypical CSS for which the 5-year survival is only 60%. Frontline therapy for HLH consists of the glucocorticoid dexamethasone (DEX) and the chemotherapeutic agent etoposide. Many patients, however, are refractory to this treatment or relapse after an initial response. Notably, many cytokines that are elevated in HLH activate the JAK/STAT pathway, and the JAK1/2 inhibitor ruxolitinib (RUX) has shown efficacy in murine HLH models and humans with refractory disease. We recently reported that cytokine-induced JAK/STAT signaling mediates DEX resistance in T cell acute lymphoblastic leukemia (T-ALL) cells, and that this could be effectively reversed by RUX. On the basis of these findings, we hypothesized that cytokine-mediated JAK/STAT signaling might similarly contribute to DEX resistance in HLH, and that RUX treatment would overcome this phenomenon. Using ex vivo assays, a murine model of HLH, and primary patient samples, we demonstrate that the hypercytokinemia of HLH reduces the apoptotic potential of CD8 T cells leading to relative DEX resistance. Upon exposure to RUX, this apoptotic potential is restored, thereby sensitizing CD8 T cells to DEX-induced apoptosis in vitro and significantly reducing tissue immunopathology and HLH disease manifestations in vivo. Our findings provide rationale for combining DEX and RUX to enhance the lymphotoxic effects of DEX and thus improve the outcomes for patients with HLH and related CSS.

Mechanisms of action of ruxolitinib in murine models of hemophagocytic lymphohistiocytosis.

Hemophagocytic lymphohistiocytosis (HLH) is an often-fatal disorder characterized by the overactivation of T cells and macrophages that excessively produce proinflammatory cytokines, including interferon-γ (IFN-γ). Previously, we reported that the JAK inhibitor ruxolitinib dampens T-cell activation and lessens inflammation in a model of HLH in which perforin-deficient (Prf1 -/-) mice are infected with lymphocytic choriomeningitis virus (LCMV). Ruxolitinib inhibits signaling downstream of IFN-γ, as well as several other JAK-dependent cytokines. As a consequence, it remained unclear whether ruxolitinib was exerting its beneficial effects in HLH by inhibiting IFN-γ signaling or by targeting signaling initiated by other proinflammatory cytokines. To address this question, we compared the effects of ruxolitinib with those obtained using an IFN-γ-neutralizing antibody (αIFN-γ) in 2 murine HLH models. In both models, ruxolitinib and αIFN-γ reduced inflammation-associated anemia, indicating that ruxolitinib operates in an IFN-γ-dependent manner to reverse this HLH manifestation. In contrast, the number and activation status of T cells and neutrophils, as well as their infiltration into tissues, were significantly reduced following treatment with ruxolitinib, but they remained unchanged or were increased following treatment with αIFN-γ. Notably, despite discontinuation of ruxolitinib, LCMV-infected Prf1 -/- mice exhibited enhanced survival compared with mice in which αIFN-γ was discontinued. This protective effect could be mimicked by transient treatment with αIFN-γ and a neutrophil-depleting antibody. Thus, ruxolitinib operates through IFN-γ-dependent and -independent mechanisms to dampen HLH by targeting the deleterious effects of T cells and neutrophils, with the latter representing an unappreciated and understudied cell type that contributes to HLH pathogenesis.

Synthesis and Biological Studies of (+)-Liquiditerpenoic Acid A (Abietopinoic Acid) and Representative Analogues: SAR Studies.

The first semisynthesis and biological profiling of the new abietane diterpenoid (+)-liquiditerpenoic acid A (abietopinoic acid) (7) along with several analogues are reported. The compounds were obtained from readily available methyl dehydroabietate (8), which was derived from (-)-abietic acid (1). Biological comparison was conducted according to the different functional groups, leading to some basic structure-activity relationships (SAR). In particular, the ferruginol and sugiol analogues 7 and 10-16 were characterized by the presence of an acetylated phenolic moiety, an oxidized C-7 as a carbonyl, and a different functional group at C-18 (methoxycarbonyl, carboxylic acid, and hydroxymethyl). The biological properties of these compounds were investigated against a panel of six representative human tumor solid cells (A549, HBL-100, HeLa, SW1573, T-47D, and WiDr), five leukemia cellular models (NALM-06, KOPN-8, SUP-B15, UoCB1, and BCR-ABL), and four Leishmania species ( L. infantum, L. donovani, L. amazonensis, and L. guyanensis). A molecular docking study pointed out some targets in these Leishmania species. In addition, the ability of the compounds to modulate GABAA receptors (α1ß2γ2s) is also reported. The combined findings indicate that these abietane diterpenoids offer a source of novel bioactive molecules with promising pharmacological properties from cheap chiral-pool building blocks.

Identification of substituted 5-membered heterocyclic compounds as potential anti-leukemic agents.

Although pediatric leukemia is generally treatable, certain leukemic subtypes face poor prognosis in the clinic suggesting new selective therapeutic agents are needed. Thus, to identify selective apoptosis inducers, a small-molecule library screening approach was conducted using an isogenic leukemic murine p185+ B-ALL cell line pair (BCR-ABL-WT and the BAX/BAK deficient BCR-ABL-DKO). Gratifyingly, the investigation revealed several compounds featuring substituted aromatic five-membered-ring heterocycles with significant activity against murine and human leukemic cellular models. The identified compounds represent potentially novel antileukemic molecular scaffolds exemplified by compounds 1, 2 and 7, which demonstrated EC50 values in the nanomolar and low micromolar range against various leukemia subtypes (SUP-B15, KOPN-8, NALM-06, UoC-B1 cellular models) and pro-apoptotic properties in solid tumor cell models (MDA-MB-231, SUM149) with ample therapeutic index in normal cells. Herein, we highlight compounds 1, 2 and 7 which promote cell death mediated by caspase 3/7 induction. Our study establishes a strategic platform for the development of potent and selective anti-leukemic agents.

The relationship between moderate alcohol consumption, depressive symptomatology, and C-reactive protein: the Health and Retirement Study.

OBJECTIVE: Moderate alcohol use has been broadly associated with health benefits among older adults, including improved mood. Aims of this study were to evaluate the relationship of moderate alcohol use and depressive symptomatology over a period of eight years, and to examine inflammation, indicated by C-reactive protein (CRP), as one mechanism by which this relationship functions. METHODS: The study included 3177 community-dwelling participants over the age of 65 in 2008 drawn from the Health and Retirement Study. Data from the 2006, 2008, 2012, and 2014 waves were used. Alcohol use was measured via self-report and was dichotomized as abstinent (0 drinks per week) and moderate (1-14 drinks per week). Inflammation was measured using CRP, which was collected using an enzyme-linked immunosorbent assay and provided in units of µg/mL. Control variables included gender, age, body mass index (BMI), and medical burden. RESULTS: A latent growth curve model with full information maximum likelihood was used, with results revealing that moderate drinkers endorsed fewer depressive symptoms at baseline and a steeper rate of change over time. Abstinent respondents' depressive symptomatology was characterized by a more linear change rate. Further, moderate drinkers had lower CRP levels suggesting that inflammation partially mediates the relationship between moderate alcohol use and depressive symptomatology. CONCLUSIONS: Moderate alcohol use predicts fewer depressive symptoms among older adults. This relationship is partially moderated by CRP and is eroded by the passage of time. Future research should identify additional mechanisms relating alcohol to positive health outcomes and less depressive symptomatology. Copyright © 2017 John Wiley & Sons, Ltd.

When Employment and Caregiving Collide: Predictors of Labor Force Participation in Prospective and Current Caregivers.

OBJECTIVE: Female caregivers often reduce time spent at work to care for aging family members, which precipitates financial hardship and other adverse outcomes. Little is known about psychosocial correlates of labor force participation (LFP) among female caregivers. The theory of planned behavior posits that social norms, attitudes, and perceived control predict intentions and volitional behaviors, but also that the compelling influence of situational variables undermines enactment of behaviors consistent with one's intentions. The objective of this study was to employ the theory of planned behavior to examine how psychosocial characteristics predict willingness to reduce LFP among prospective caregivers and actual LFP reduction among active caregivers. METHODS: Subsamples of 165 female prospective caregivers and 97 active female caregivers responded to a survey assessing filial beliefs and LFP. RESULTS: Filial obligation and caregiver preparedness predicted intentions to reduce LFP among prospective caregivers, but did not predict actual reduction in LFP in active caregivers. Weekly care demands exceeding 20 hours emerged as the sole correlate of LFP among active caregivers. CONCLUSIONS: Domains of the theory of planned behavior predict LFP intentions, but LFP decisions are subject to external pressures, specifically, time demands of the caregiving relationship. Prospective caregivers may benefit from proactive interventions aimed at reducing conflict between situational demands and filial beliefs.

Autotransfusion in dogs using a 2-syringe technique.

OBJECTIVE: To describe the successful use of an autotransfusion technique utilizing 2 syringes in 4 dogs. CASE SERIES SUMMARY: All 4 dogs in this series had a hemoabdomen and subsequent hypovolemic shock. During surgery blood was collected from the abdominal cavity by the surgeon and passed to an assistant. The blood was then transferred to a second syringe for direct IV administration. The blood was passed through an inline blood filter prior to reaching the patient. Given the transfusion volume and administration time frame, 3 cases were classified as a massive transfusion. All 4 dogs survived the transfusion, were discharged within 3 days of surgery/transfusion and no complications were noted. NEW OR UNIQUE INFORMATION PROVIDED: This case series describes a relatively simple method of performing an autotransfuion in patients with hemoabdomen and hypovolemic shock.

Prepared to care: adult attachment and filial obligation.

OBJECTIVES: Past work found that close adult attachment dimension scores predict caregiver preparedness. Theory and past research suggests filial obligation (FO) may mediate the relationship between attachment and caregiver preparedness. The goal of this study was to test that hypothesis. METHOD: The sample, collected using Mechanical Turk, included 165 women between the ages of 45 and 65 years who were not providing care to an aging parent.  Participants were reimbursed $0.75 for completing an online survey assessing response validity, dimensions of adult attachment, depressive symptomatology, FO, and caregiver preparedness, among other variables. The mean participant age was 52.2 years (SD = 5.5). RESULTS: The sample was predominantly White/Caucasian (82.6%), and Black/African American (7.3%).  With respect to education, 14.5% completed high school or general education development (GED), 32.7% completed some college, 36.4% completed a Bachelor's degree, and 15.7% completed graduate degrees. The three attachment dimensions (close, dependent, and anxious) and FO were all significantly inner-correlated.  Stepwise multiple regression analyses found that FO fully mediates the relationship between close attachment and caregiver preparedness, even after controlling for age, education, income, depression, and birth order. CONCLUSION: The primary finding is that FO mediates the relationship between close attachment style and caregiver preparedness among prospective caregivers. This suggests that individual differences in attachment style among prospective caregivers indirectly predict preparedness for future caregiving through FO, suggesting a mechanism relating attachment style and preparation for future care.