Single Ascending and Multiple-Dose Trial of Zerlasiran, a Short Interfering RNA Targeting Lipoprotein(a): A Randomized Clinical Trial.

Importance: Lipoprotein(a) is a causal risk factor for atherosclerotic cardiovascular disease (ASCVD) and calcific aortic stenosis, with no pharmacological treatments approved by regulatory authorities. Objectives: To assess the safety and tolerability of zerlasiran, a short interfering RNA targeting hepatic synthesis of apolipoprotein(a), and effects on serum concentrations of lipoprotein(a). Design, Setting, and Participants: Single- and multiple-dose study in healthy participants and patients with stable ASCVD, respectively, with lipoprotein(a) serum concentrations greater than 150 nmol/L, conducted at 7 research sites in the US, the Netherlands, UK, and Australia between November 18, 2020, and February 8, 2023, with last follow-up on August 23, 2023. Interventions: Participants were randomized to receive (1) a single subcutaneous dose of placebo (n = 8), zerlasiran 300 mg (n = 6) or 600 mg (n = 6); or (2) 2 doses of placebo (n = 9), zerlasiran 200 mg (n = 9) at a 4-week interval or 300 mg (n = 9) or 450 mg (n = 9) at an 8-week interval. Main Outcomes Measures: The primary outcome was safety and tolerability. Secondary outcomes included serum levels of zerlasiran and effects on lipoprotein(a) serum concentrations. Results: Among 37 patients in the multiple-dose group (mean age, 56 [SD, 10.4] years; 15 [42%] women), 36 completed the trial. Among 14 participants with extended follow-up after single doses, 13 completed the trial. There were no serious adverse events. Median baseline lipoprotein(a) concentrations in the multiple-dose group were 288 (IQR, 199-352) nmol/L. Median changes in lipoprotein(a) concentration at 365 days after single doses were 14% (IQR, 13% to 15%) for the placebo group, -30% (IQR, -51% to -18%) for the 300 mg of zerlasiran group, and -29% (IQR, -39% to -7%) for the 600-mg dose group. After 2 doses, maximal median changes in lipoprotein(a) concentration were 19 (IQR, -17 to 28) nmol/L for the placebo group, -258 (IQR, -289 to -188) nmol/L for the 200 mg of zerlasiran group, -310 (IQR, -368 to -274) nmol/L for the 300-mg dose group, and -242 (IQR, -343 to -182) nmol/L for the 450-mg dose group, with maximal median percent change of 7% (IQR, -4% to 21%), -97% (IQR, -98% to -95%), -98% (IQR, -99% to -97%), and -99% (IQR, -99% to -98%), respectively, attenuating to 0.3% (IQR, -2% to 21%), -60% (IQR, -71% to -40%), -90% (IQR, -91% to -74%), and -89% (IQR, -91% to -76%) 201 days after administration. Conclusions: Zerlasiran was well tolerated and reduced lipoprotein(a) concentrations with infrequent administration. Trial Registration: ClinicalTrials.gov Identifier: NCT04606602.

Semiautomated approach focused on new genomic information results in time and effort-efficient reannotation of negative exome data.

Most rare disease patients (75-50%) undergoing genomic sequencing remain unsolved, often due to lack of information about variants identified. Data review over time can leverage novel information regarding disease-causing variants and genes, increasing this diagnostic yield. However, time and resource constraints have limited reanalysis of genetic data in clinical laboratories setting. We developed RENEW, (REannotation of NEgative WES/WGS) an automated reannotation procedure that uses relevant new information in on-line genomic databases to enable rapid review of genomic findings. We tested RENEW in an unselected cohort of 1066 undiagnosed cases with a broad spectrum of phenotypes from the Mayo Clinic Center for Individualized Medicine using new information in ClinVar, HGMD and OMIM between the date of previous analysis/testing and April of 2022. 5741 variants prioritized by RENEW were rapidly reviewed by variant interpretation specialists. Mean analysis time was approximately 20 s per variant (32 h total time). Reviewed cases were classified as: 879 (93.0%) undiagnosed, 63 (6.6%) putatively diagnosed, and 4 (0.4%) definitively diagnosed. New strategies are needed to enable efficient review of genomic findings in unsolved cases. We report on a fast and practical approach to address this need and improve overall diagnostic success in patient testing through a recurrent reannotation process.

Revisiting Stl's thoughts: formalizing the ictericus group in Euschistus (Euschistus) (Hemiptera: Heteroptera: Pentatomidae).

Euschistus Dallas is distributed throughout the Western Hemisphere and is composed of 67 species in three subgenera: Euschistomorphus Jensen-Haarup, Lycipta Stl, and the nominate subgenus. Euschistus (Euschistus) includes several economically important species. Due to the lack of a phylogenetic analysis for the subgenus Euschistus, its groupings are based primarily on male genitalia. In this paper, the following Nearctic taxa that were first grouped together by Stl (1872), namely E. ictericus (Linnaeus), E. variolarius (Palisot de Beauvois), E. servus servus (Say), E. servus euschistoides (Vollenhoven), and E. tristigmus tristigmus (Say), are reviewed and redescribed. Several other similar, and probably related, taxa are also included in this study: E. tristigmus luridus (Dallas), E. inflatus Van Duzee, and E. latimarginatus Zimmer. Morphological patterns pertaining to the internal and external male and female genitalia allowed us to formalize the ictericus group to include the above-mentioned species (except E. tristigmus). The morphological overlap between the subspecies and sympatric areas of distribution have led us to consider E. servus euschistoides as a junior synonym of E. servus servus, and to corroborate the synonymy first proposed by Uhler (1861) of E. tristigmus luridus under E. tristigmus tristigmus.

Preclinical Toxicological Assessment of A Novel siRNA, SLN360, Targeting Elevated Lipoprotein (a) in Cardiovascular Disease.

SLN360 is a liver-targeted N-acetyl galactosamine (GalNAc)-conjugated small interfering RNA (siRNA) with a promising profile for addressing lipoprotein (a)-related cardiovascular risk. Here, we describe the findings from key preclinical safety studies. In vitro, SLN360 specifically reduced LPA expression in primary human hepatocytes with no relevant off-target effects. In rats, 10 mg/kg subcutaneous SLN360 was distributed specifically to the liver and kidney (peak 126 or 246 mg/g tissue at 6 h, respectively), with <1% of peak liver levels observed in all other tested organs. In vitro, no genotoxicity and no effect on human Ether-a-go-go Related Gene currents or proinflammatory cytokine production was observed, whereas in vivo, no SLN360-specific antibodies were detected in rabbit serum. In rat and nonhuman primate 29-day toxicology studies, SLN360 was well tolerated at all doses. In both species, known GalNAc-conjugated siRNA-induced microscopic changes were observed in the kidney and liver, with small increases in alanine aminotransferase and alkaline phosphatase observed in the high dose rats. Findings were in line with previously described siRNA-GalNAc platform-related effects and all observations were reversible and considered nonadverse. In cynomolgus monkeys, liver LPA messenger RNA and serum lipoprotein (a) were significantly reduced at day 30 and after an 8-week recovery period. No dose-related changes in safety assessment endpoints were noted. No SLN360-induced cytokine production, complement activation, or micronucleus formation was observed in vivo. The toxicological profile of SLN360 presented here is restricted to known GalNAc siRNA effects and no other toxicity associated with SLN360 has been noted. The preclinical profile of SLN360 confirmed suitability for entry into clinical studies.

Pre-clinical assessment of SLN360, a novel siRNA targeting LPA, developed to address elevated lipoprotein (a) in cardiovascular disease.

BACKGROUND AND AIMS: The LPA gene encodes apolipoprotein (a), a key component of Lp(a), a potent risk factor for cardiovascular disease with no specific pharmacotherapy. Here we describe the pharmacological data for SLN360, a GalNAc-conjugated siRNA targeting LPA, designed to address this unmet medical need. METHODS: SLN360 was tested in vitro for LPA knockdown in primary hepatocytes. Healthy cynomolgus monkeys received single or multiple subcutaneous doses of the SLN360 sequence ranging from 0.1 to 9.0 mg/kg to determine the pharmacokinetic and pharmacodynamic effects. Liver mRNA and serum biomarker analyses were performed. RESULTS: In vitro, the SLN360 sequence potently reduces LPA mRNA in primary cynomolgus and human hepatocytes, while no effect was observed on the expression of APOB or PLG. In vivo, SLN360 exposure peaks 2 h after subcutaneous injection with near full elimination by 24 h. Specific LPA mRNA reduction (up to 91% 2 weeks after dosing) was observed with only the 3 mg/kg group showing appreciable return to baseline (40%). No consistent dose- or time-dependent effect on the expression of APOB, PLG or a panel of sensitive markers of liver lipid accumulation was observed. Potent (up to 95%) and long lasting (≥9 weeks) serum Lp(a) reduction was observed, peaking in all active groups at day 21. The minimally effective dose was determined to be 0.3 mg/kg with an ED50 of 0.6 mg/kg. CONCLUSIONS: SLN360 induces a sustained reduction in serum Lp(a) levels in cynomolgus monkeys following subcutaneous dosing. SLN360 has potential to address the unmet need of Lp(a) reduction in cardiovascular diseases.

Treatment and prevention of lipoprotein(a)-mediated cardiovascular disease: the emerging potential of RNA interference therapeutics.

Lipid- and lipoprotein-modifying therapies have expanded substantially in the last 25 years, resulting in reduction in the incidence of major adverse cardiovascular events. However, no specific lipoprotein(a) [Lp(a)]-targeting therapy has yet been shown to reduce cardiovascular disease risk. Many epidemiological and genetic studies have demonstrated that Lp(a) is an important genetically determined causal risk factor for coronary heart disease, aortic valve disease, stroke, heart failure, and peripheral vascular disease. Accordingly, the need for specific Lp(a)-lowering therapy has become a major public health priority. Approximately 20% of the global population (1.4 billion people) have elevated levels of Lp(a) associated with higher cardiovascular risk, though the threshold for determining 'high risk' is debated. Traditional lifestyle approaches to cardiovascular risk reduction are ineffective at lowering Lp(a). To address a lifelong risk factor unmodifiable by non-pharmacological means, Lp(a)-lowering therapy needs to be safe, highly effective, and tolerable for a patient population who will likely require several decades of treatment. N-acetylgalactosamine-conjugated gene silencing therapeutics, such as small interfering RNA (siRNA) and antisense oligonucleotide targeting LPA, are ideally suited for this application, offering a highly tissue- and target transcript-specific approach with the potential for safe and durable Lp(a) lowering with as few as three or four doses per year. In this review, we evaluate the causal role of Lp(a) across the cardiovascular disease spectrum, examine the role of established lipid-modifying therapies in lowering Lp(a), and focus on the anticipated role for siRNA therapeutics in treating and preventing Lp(a)-related disease.

A distributional synopsis of the Pentatomidae (Heteroptera) north of Mexico, including new state and provincial records.

The species of Pentatomidae known to occur north of Mexico, comprising 223 species in 68 genera, are enumerated with taxonomic notes and updated and annotated distributions. Included in this update are 126 new state records reported for 62 pentatomid species in 30 genera. The copious annotations in these distributions and attendant bibliography serve as an extensive compilation of overlooked references that might contain distributional records for other insect, especially heteropteran, species.

Finding Padaeus bovillus (Hemiptera: Pentatomidae): A Phylogenetic Placement and the Description of Its Sister Species.

The pentatomids (Hemiptera: Heteroptera) are the third most speciose family within the Heteroptera or the true bugs. The family occurs worldwide and comprises around five thousand valid species within 950 genera. Padaeus Stål belongs to a complex of other genera of Carpocorini Mulsant and Rey related to Euschistus Dallas. These genera present similarities in color, size, and shape, and share common features. However, among its four congeneric species, Padaeus bovillus Distant has been highlighted as an outlier by the posterior margins of the bucculae evanescent, while its congeneric species present posterior margins of the bucculae lobed. Thus, herein we redescribe P. bovillus and present a hypothesis regarding its phylogenetic placement within the Carpocorini. Furthermore, a new species similar to P. bovillus is described. Four molecular markers (COI, CytB, 16S, and 28S) plus 86 morphological characters were used to infer the phylogeny under Maximum likelihood and Bayesian inference. For the descriptions, we measured 16 morphometric parameters and dissected the genitalic structures. We also include illustrations of the habitus, internal and external genitalic structures, and provide distribution maps. Mitripus seclusus sp. n. Bianchi, Krein, Rider, and Grazia is recovered as the sister species to Mitripus bovillus comb. n., and both within Mitripus Rolston. Among other shared characters, species of Mitripus have the femora unarmed, they have a macula near the apex of the radial vein, and the mandibular plates tapering apically. Mitripus bovillus comb. n. and Mitripus seclusus sp. n. have the posterior margin of the pygophore projecting as a spine, a unique pattern within the genus. According to our results, Mitripus including M. bovillus comb. n. and Mitripus seclusus sp. n. now includes five species.

emColimacoris/em emjoceliae/em, sp. nov. (Hemiptera: Miridae: Deraeocorinae: Hyaliodini): A new species from Panama, with a Key to the species of Colimacoris.

The new Neotropical hyaliodine Colimacoris joceliae Kim Jung, sp. nov., is described from Panama. Morphological information, including a description and diagnosis of the new species is presented with photographs. A key for the identification of the species of Colimacoris is also provided.

Two new species of Tingidae (Hemiptera: Heteroptera) from Panama.

Two new species of Tingidae are described from Panama: Acanthotingis deltoides sp. nov. and Stephanitis joceliae sp. nov. Descriptions and diagnosis for both Acanthotingis Monte and Stephanitis Stål are provided, and a key is given for the separation of the two known species of Acanthotingis. This is the first record of both genera from Panama. A checklist of all Neotropical species of both genera is presented.

New genus and new species of Pentatomidae from Borneo (Hemiptera: Heteroptera).

A new genus, Graziasternum gen. nov., with two new species, Graziasternum graziae sp. nov. and Graziasternum joceliae sp. nov. are described. According to external thoracic and genital structures, this new genus is considered to be the closest extant relative known to the genus Placosternum Amyot Serville. Discussion of its possible phylogenetic position based on structure comparison, with the discussion on the importance of the characteristics of the thoracic sterna, the genital structures with other genera or groups of genera that share similar structures is given. Additionally, the two new species are keyed and illustrated.

emGraziacrophyma/em gen. nov., a new genus of South American Acanthosomatidae (Hemiptera: Heteroptera).

The genus Acrophyma Bergroth currently contains two species, the Andean A. cumingii (Westwood) and the Neotropical A. bicallosa (Stål). The examination of types and new material revealed that among the Neotropical species the Brazilian population differs from the Colombian, and the first one belongs to a new species in this group. A morphological analysis of the genus indicated the Neotropical species are different from the Andean species. According to the current most used generic characters in acanthosomatids, Neotropical species should be placed in a new genus. This is also supported by biogeographical relationships. New distributional records for the species of this group are provided.

Development and Application of Nanoparticle-Nanopolymer Composite Spheres for the Study of Environmental Processes.

Plastics have long been an environmental contaminant of concern as both large-scale plastic debris and as micro- and nano-plastics with demonstrated wide-scale ubiquity. Research in the past decade has focused on the potential toxicological risks posed by microplastics, as well as their unique fate and transport brought on by their colloidal nature. These efforts have been slowed by the lack of analytical techniques with sufficient sensitivity and selectivity to adequately detect and characterize these contaminants in environmental and biological matrices. To improve analytical analyses, microplastic tracers are developed with recognizable isotopic, metallic, or fluorescent signatures capable of being identified amidst a complex background. Here we describe the synthesis, characterization, and application of a novel synthetic copolymer nanoplastic based on polystyrene (PS) and poly(2-vinylpyridine) (P2VP) intercalated with gold, platinum or palladium nanoparticles that can be capped with different polymeric shells meant to mimic the intended microplastic. In this work, particles with PS and polymethylmethacrylate (PMMA) shells are used to examine the behavior of microplastic particles in estuarine sediment and coastal waters. The micro- and nanoplastic tracers, with sizes between 300 and 500 nm in diameter, were characterized using multiple physical, chemical, and colloidal analysis techniques. The metallic signatures of the tracers allow for quantification by both bulk and single-particle inductively-coupled plasma mass spectrometry (ICP-MS and spICP-MS, respectively). As a demonstration of environmental applicability, the tracers were equilibrated with sediment collected from Bellingham Bay, WA, United States to determine the degree to which microplastics bind and sink in an estuary based of grain size and organic carbon parameters. In these experiments, between 80 and 95% of particles were found to associate with the sediment, demonstrative of estuaries being a major anticipated sink for these contaminants. These materials show considerable promise in their versatility, potential for multiplexing, and utility in studying micro- and nano-plastic transport in real-world environments.

Taxonomic review of Sibaria Stål, 1872 (Heteroptera: Pentatomidae: Pentatominae: Carpocorini), with description of a new species.

Sibaria Stål is a genus of Pentatomidae occurring exclusively in the Neotropical region. Since its description, little information has been added to the knowledge of the taxon. The hypothesis of monophyly of Sibaria was recently supported by a phylogenetic analysis based on morphological and molecular data. The purpose of this study is to provide a review of Sibaria, redescribe the valid species, and describe a new species. The genus includes four species: S. andicola Breddin, S. armata (Dallas), S. englemani Rolston, and S. amazonica sp. nov.. Diagnoses, macrophotographs (habitus, external and internal genitalia), an identification key, and distribution records of the species are also provided.

Central IRAK-4 kinase inhibition for the treatment of pain following nerve injury in rats.

There is ample evidence for the role of the immune system in developing chronic pain following peripheral nerve injury. Especially Toll-like receptors (TLRs) and their associated signaling components and pro-inflammatory cytokines such as IL-1ß, induced after injury, are involved in nociceptive processes and believed to contribute to the manifestation of chronic neuropathic pain states. Whereas the inhibition of the kinase function of IRAK-4, a central kinase downstream of TLRs and IL-1 receptors (IL-1Rs), seems efficacious in various chronic inflammatory and autoimmune models, it's role in regulating chronic neuropathic pain remained elusive to date. Here, we examined whether pharmacological inhibition of IRAK-4 kinase activity using PF-06650833 and BMS-986147, two clinical-stage kinase inhibitors, is effective for controlling persistent pain following nerve injury. Both inhibitors potently inhibited TLR-triggered cytokine release in human peripheral blood mononuclear cell (PBMC) as well as human and rat whole blood cultures. BMS-986147 showing favorable pharmacokinetic (PK) properties, significantly inhibited R848-triggered plasma TNF levels in a rat in vivo cytokine release model after single oral dosing. However, BMS-986147 dose dependently reversed cold allodynia in a rat chronic constriction injury (CCI) model following intrathecal administration only, supporting the notion that central neuro-immune modulation is beneficial for treating chronic neuropathic pain. Although both inhibitors were efficacious in inhibiting IL-1ß- or TLR-triggered cytokine release in rat dorsal root ganglion cultures, only partial efficacy was reached in IL-1ß-stimulated human glial cultures indicating that inhibiting IRAK-4́'s kinase function might be partially dispensable for human IL-1ß driven neuroinflammation. Overall, our data demonstrate that IRAK-4 inhibitors could provide therapeutic benefit in chronic pain following nerve injury, and the central driver for efficacy in the neuropathic pain model as well as potential side effects of centrally available IRAK-4 inhibitors warrant further investigation to develop effective analgesia for patients in high unmet medical need.

FPR-1 is an important regulator of neutrophil recruitment and a tissue-specific driver of pulmonary fibrosis.

Neutrophils are the most abundant inflammatory cells at the earliest stages of wound healing and play important roles in wound repair and fibrosis. Formyl peptide receptor 1 (FPR-1) is abundantly expressed on neutrophils and has been shown to regulate their function, yet the importance of FPR-1 in fibrosis remains ill defined. FPR-1-deficient (fpr1-/-) mice were protected from bleomycin-induced pulmonary fibrosis but developed renal and hepatic fibrosis normally. Mechanistically, we observed a failure to effectively recruit neutrophils to the lungs of fpr1-/- mice, whereas neutrophil recruitment was unaffected in the liver and kidney. Using an adoptive transfer model we demonstrated that the defect in neutrophil recruitment to the lung was intrinsic to the fpr1-/- neutrophils, as C57BL/6 neutrophils were recruited normally to the damaged lung in fpr1-/- mice. Finally, C57BL/6 mice in which neutrophils had been depleted were protected from pulmonary fibrosis. In conclusion, FPR-1 and FPR-1 ligands are required for effective neutrophil recruitment to the damaged lung. Failure to recruit neutrophils or depletion of neutrophils protects from pulmonary fibrosis.

A new genus and new species of Aeptini (Hemiptera: Heteroptera: Pentatomidae: Pentatominae) from Australia.

The aeptine Mariomella singularis gen. et sp. nov. is described and illustrated from Western Australia. With this new addition, the Aeptini diversity increases to 21 species in nine genera.

Haplosaurus computes protein haplotypes for use in precision drug design.

Selecting the most appropriate protein sequences is critical for precision drug design. Here we describe Haplosaurus, a bioinformatic tool for computation of protein haplotypes. Haplosaurus computes protein haplotypes from pre-existing chromosomally-phased genomic variation data. Integration into the Ensembl resource provides rapid and detailed protein haplotypes retrieval. Using Haplosaurus, we build a database of unique protein haplotypes from the 1000 Genomes dataset reflecting real-world protein sequence variability and their prevalence. For one in seven genes, their most common protein haplotype differs from the reference sequence and a similar number differs on their most common haplotype between human populations. Three case studies show how knowledge of the range of commonly encountered protein forms predicted in populations leads to insights into therapeutic efficacy. Haplosaurus and its associated database is expected to find broad applications in many disciplines using protein sequences and particularly impactful for therapeutics design.

Kaytuesso flavolateralis gen. and sp. nov., a new monotypic genus of Oncomerinae (Hemiptera: Heteroptera: Tessaratomidae) from Papua New Guinea.

Kaytuesso flavolateralis gen. and sp. nov. is described from Papua New Guinea. The new genus is remarkable in that the only included species is one of the smallest species known within the Oncomerinae. The relationships among Kaytuesso and the related genera Agapophyta Guerin, Erga Walker and Musgraveia Leston and Scudder are discussed.

Review of the genus Menestheus Stål, 1868 (Hemiptera: Heteroptera: Pentatomidae).

The Aeptini (Pentatomidae: Pentatominae) genus Menestheus Stål, 1868, is redescribed. The original misidentification of the type species for Menestheus is corrected by action of first reviser herein by establishing Paramenestheus nercivus non Dallas, 1851 sensu Stal (1868) = Menestheus cuneatus Distant, 1899 as the type species. Menestheus mcphersoni sp. nov. is described and illustrated. Characters separating the two species are discussed.