Monitoring Pharmacologically Induced Immunosuppression by Immune Repertoire Sequencing to Detect Acute Allograft Rejection in Heart Transplant Patients: A Proof-of-Concept Diagnostic Accuracy Study

Background

It remains difficult to predict and to measure the efficacy of pharmacological immunosuppression. We hypothesized that measuring the B-cell repertoire would enable assessment of the overall level of immunosuppression after heart transplantation.

Methods and Findings

In this proof-of-concept study, we implemented a molecular-barcode-based immune repertoire sequencing assay that sensitively and accurately measures the isotype and clonal composition of the circulating B cell repertoire. We used this assay to measure the temporal response of the B cell repertoire to immunosuppression after heart transplantation. We selected a subset of 12 participants from a larger prospective cohort study (ClinicalTrials.gov NCT01985412) that is ongoing at Stanford Medical Center and for which enrollment started in March 2010. This subset of 12 participants was selected to represent post-heart-transplant events, with and without acute rejection (six participants with moderate-to-severe rejection and six without). We analyzed 130 samples from these patients, with an average follow-up period of 15 mo. Immune repertoire sequencing enables the measurement of a patient’s net state of immunosuppression (correlation with tacrolimus level, r = −0.867, 95% CI −0.968 to −0.523, p = 0.0014), as well as the diagnosis of acute allograft rejection, which is preceded by increased immune activity with a sensitivity of 71.4% (95% CI 30.3% to 94.9%) and a specificity of 82.0% (95% CI 72.1% to 89.1%) (cell-free donor-derived DNA as noninvasive gold standard). To illustrate the potential of immune repertoire sequencing to monitor atypical post-transplant trajectories, we analyzed two more patients, one with chronic infections and one with amyloidosis. A larger, prospective study will be needed to validate the power of immune repertoire sequencing to predict rejection events, as this proof-of-concept study is limited to a small number of patients who were selected based on several criteria including the availability of a large number of samples and the absence or presence of rejection events.

Conclusions

If confirmed in larger, prospective studies, the method described here has potential applications in the tailored management of post-transplant immunosuppression and, more broadly, as a method for assessing the overall activity of the immune system.     

Piscivorous Bird Use of Aquaculture and Natural Water Bodies in Mississippi

Double-crested cormorants (Phalacrocorax auritus) and great egrets (Ardea alba) have an extensive history of human-wildlife conflict with the aquaculture industry of western Mississippi, USA, due to their depredation of cultured catfish (Ictalurus spp.). Although aquaculture is abundant, western Mississippi also contains naturally occurring water bodies that offer alternative forage opportunities to these species. How cormorants or egrets distribute themselves among these 2 foraging options is unknown, but it has been generally assumed each species uses aquaculture disproportionately more because of the high density of available prey. To test this assumption, we surveyed these species on aquaculture and naturally occurring water bodies using aerial surveys from October through April of 2015–2016, 2016–2017, and 2017–2018. We modeled the proportion of each species on aquaculture as a function of year, date, and weather-related variables using quasi-binomial generalized linear models. Egrets used aquaculture consistently more than what was proportionally available to them and use was not influenced by any of the variables we measured. Proportional use of aquaculture by cormorants was lowest during October through January but steadily increased through April, indicating a distribution shift toward aquaculture in the months immediately prior to their migration. The highest proportional use of aquaculture by cormorants occurred in 2016, a year when lethal control measures were not allowed against cormorants. Conversely, the least proportion of cormorants on aquaculture was in 2015 when cormorants could be lethally controlled under authority of an Aquaculture Depredation Order. This trend highlights the potential influence of changes in mortality risk, caused by changes in policy regarding lethal take of cormorants, on cormorant distribution between foraging options. © 2020 The Wildlife Society.     

Dual inhibitors of inosine monophosphate dehydrogenase and histone deacetylase based on a cinnamic hydroxamic acid core structure

Small molecules that act on multiple biological targets have been proposed to combat the drug resistance commonly observed for cancer chemotherapy. By combining the structural features of known inhibitors of inosine monophosphate dehydrogense (IMPDH) and histone deacetylase (HDAC), dual inhibitors of IMPDH and HDAC based on the scaffold of cinnamic hydroxamic acid (CHA) have been designed, synthesized, and evaluated in biological assays. Key features, including the linker length, linker functionality, substitution position, and interacting groups, have been explored. Their individual contribution to the inhibitory activities against human IMPDH1 and IMPDH2 as well as HDAC has been assessed.     

Synthesis of orthogonally reactive FK506 derivatives via olefin cross metathesis

Chemical inducers of dimerization (CIDs) are employed in a wide range of biological applications to control protein localization, modulate protein–protein interactions and improve drug lifetimes. These bifunctional chemical probes are assembled from two synthetic modules, which each provide affinity for a distinct protein target. FK506 and its derivatives are often employed as modules in the syntheses of these bifunctional constructs, owing to the abundance and favorable distribution of their target, FK506-binding protein (FKBP). However, the structural complexity of FK506 necessitates multi-step syntheses and/or multiple protection–deprotection schemes prior to installation into CIDs. In this work, we describe an efficient, one-step synthesis of FK506 derivatives through a selective, microwave-accelerated, cross metathesis diversification step of the C39 terminal alkene. Using this approach, FK506 is modified with an array of functional groups, including primary amines and carboxylic acids, which make the resulting derivatives suitable for the modular assembly of CIDs. To illustrate this idea, we report the synthesis of a heterobifunctional HIV protease inhibitor.