Background: Motixafortide is a novel, high affinity CXCR4 antagonist with extended receptor occupancy that facilitates the migration of hematopoietic stem and progenitor cells from the bone marrow ...into the peripheral blood in preparation for autologous stem cell transplantation (ASCT). In prior pre-clinical and clinical studies, motixafortide was associated with transient hypersensitivity and injection site reactions. Strategies to mitigate such adverse events (AEs) commonly include agents with antihistamine activity. The Phase 3 GENESIS trial demonstrated that 88.8% of Multiple Myeloma patients mobilized ≥6x10 6 CD34+ cells/kg (local labs) in 1 leukapheresis procedure (LP) following the administration of 1 dose of motixafortide in combination with GCSF. Per initial protocol, premedication with an antihistamine H1 blocker prior to motixafortide administration was provided to all patients. However, during the course of the GENESIS study a protocol amendment was implemented to include premedication with an antihistamine H1 blocker, an H2 blocker, a leukotriene inhibitor and acetaminophen prior to motixafortide administration. Here we report the safety findings in a cohort of patients at who received single drug premedication with an H1 antihistamine prior to the protocol amendment and those who received the updated combination pre-medication regimen following protocol amendment. Methods:The GENESIS trial was a multicenter, randomized, double-blind, placebo-controlled study in 122 transplant-eligible multiple myeloma patients, as previously described (PMID: 37069359). Included in this analysis are 46 patients enrolled at the Washington University in St. Louis; 32 patients were randomized to the motixafortide treatment arm and 14 patients were randomized to the placebo arm. Among motixafortide-treated patients, 22 received a single-agent H1 blocker pre-medication and 10 received a combination pre-medication regimen comprising of an H1 blocker, an H2 blocker, a leukotriene inhibitor and acetaminophen. All pre-medications were administered within 1 hour prior to motixafortide administration. Hypersensitivity AEs included all events coded to the MedDRA terms of urticaria, pruritus, flushing, erythema, rash, hypersensitivity or anaphylaxis. Injection site reactions included all events coded to the MedDRA High Level Term of Injection Site Reaction. All AEs were graded using CTCAE criteria and were recorded up to 30 days from the last dose of motixafortide. Results: Overall, motixafortide was safe and well tolerated with no episodes of anaphylaxis, no Grade 4 AEs and no deaths. The most commonly observed AEs included transient hypersensitivity and injection site reactions. The frequency of motixafortide-associated hypersensitivity AEs of any grade was reduced from 90.9% with single-agent H1 blocker pre-medication to 20% with the combination pre-medication regimen, similar to the 20-25% rate of hypersensitivity AEs reported in the placebo + GCSF cohort (Table 1). In addition, all hypersensitivity AEs that occurred following combination pre-medication were graded mild in severity (Figure 1). Meanwhile the frequency of injection site AEs of any grade was reduced from 95.5% with single-agent H1 blocker pre-medication to 80.0% with the combination pre-medication regimen (Table 1). Furthermore, a notable improvement in severity of injection site AEs was observed, with moderate-severe injection site AEs reduced from 27% with single-agent H1 blocker pre-medication to 0% with the combination pre-medication regimen (Figure 1). Conclusions:Motixafortide was safe and well tolerated with no episodes of anaphylaxis, no Grade 4 AEs and no deaths. Following implementation of a protocol amendment using combination pre-medication with an antihistamine H1 blocker, an H2 blocker, a leukotriene inhibitor and acetaminophen, the frequency and severity of hypersensitivity and injection site AEs associated with motixafortide were markedly improved.
Autologous hematopoietic stem cell transplantation (ASCT) improves survival in multiple myeloma (MM). However, many individuals are unable to collect optimal CD34
hematopoietic stem and progenitor ...cell (HSPC) numbers with granulocyte colony-stimulating factor (G-CSF) mobilization. Motixafortide is a novel cyclic-peptide CXCR4 inhibitor with extended in vivo activity. The GENESIS trial was a prospective, phase 3, double-blind, placebo-controlled, multicenter study with the objective of assessing the superiority of motixafortide + G-CSF over placebo + G-CSF to mobilize HSPCs for ASCT in MM. The primary endpoint was the proportion of patients collecting ≥6 × 10
CD34
cells kg
within two apheresis procedures; the secondary endpoint was to achieve this goal in one apheresis. A total of 122 adult patients with MM undergoing ASCT were enrolled at 18 sites across five countries and randomized (2:1) to motixafortide + G-CSF or placebo + G-CSF for HSPC mobilization. Motixafortide + G-CSF enabled 92.5% to successfully meet the primary endpoint versus 26.2% with placebo + G-CSF (odds ratio (OR) 53.3, 95% confidence interval (CI) 14.12-201.33, P < 0.0001). Motixafortide + G-CSF also enabled 88.8% to meet the secondary endpoint versus 9.5% with placebo + G-CSF (OR 118.0, 95% CI 25.36-549.35, P < 0.0001). Motixafortide + G-CSF was safe and well tolerated, with the most common treatment-emergent adverse events observed being transient, grade 1/2 injection site reactions (pain, 50%; erythema, 27.5%; pruritis, 21.3%). In conclusion, motixafortide + G-CSF mobilized significantly greater CD34
HSPC numbers within two apheresis procedures versus placebo + G-CSF while preferentially mobilizing increased numbers of immunophenotypically and transcriptionally primitive HSPCs. Trial Registration: ClinicalTrials.gov , NCT03246529.
Background: Autologous stem cell transplantation (ASCT) in multiple myeloma (MM) has been shown to improve survival compared to conventional chemotherapy alone. However, the ability to perform ASCT ...relies, in part, on collecting a sufficient number (#) of CD34+ hematopoietic stem cells (HSCs), typically from peripheral blood. The ideal HSC mobilization regimen would enable collection of optimal #s of HSCs (5-6x10 6 CD34+ cells/kg) within the minimum # of apheresis sessions possible. Yet, despite currently available G-CSF (G) based mobilization regimens and multiple apheresis days, many remain unable to collect optimal #s of HSCs. Motixafortide (M) is a novel CXCR4 inhibitor, with high affinity (IC 50 0.54-4.5 nM) and long receptor occupancy (>48 hours).
Methods: In this prospective, phase 3, double blind, placebo controlled, multicenter trial, 122 patients were randomized (2:1) to receive either M+G or placebo (P)+G for HSC mobilization prior to ASCT for MM. All patients received G (10 mcg/kg) on days 1-5 (and 6-8, if needed). Patients received either M (1.25 mg/kg, subcutaneous injection) or P on day 4 (and 6, if needed). Apheresis began day 5, with the primary (PEP) and secondary (SEP) endpoints of collecting ≥6x10 6 CD34+ cells/kg in up to 2 apheresis days or 1 day, respectively. Apheresis continued on days 6-8 if needed. Total CD34+ cells/kg were analyzed on site to determine if patients mobilized to the goal and all samples were subsequently sent for assessment by central laboratory. Patients that did not collect ≥2x10 6 CD34+ cells/kg by day 8 proceeded to rescue mobilization. The # of CD34+ cells infused was determined independently by each investigator according to local practice (minimum ≥2x10 6 CD34+ cells/kg). Analyses of the PEP/SEPs were performed on an intent-to-treat basis.
Results: Demographics between the 2 treatment arms were similar. Mobilization with M+G resulted in 92.5% of patients collecting ≥6x10 6 CD34+ cells/kg within 2 apheresis days vs 26.2% with P+G (Odds Ratio (OR) 53.3, 95% CI 14.12-201.33, p<0.0001). Furthermore, 88.8% of patients with M+G collected ≥6x10 6 CD34+ cells/kg in 1 apheresis day vs 9.5% with P+G (OR 118.0, 95% CI 25.36-549.35, p<0.0001); and 96.3% with M+G collected ≥2x10 6 CD34+ cells/kg within 1 apheresis day vs 64.3% with P+G (OR 18.9, 95% CI 4.47-80.04, p<0.0001). The PEP and SEPs were confirmed as statistically significant by central laboratory (all respective p-values <0.0001). The median # of HSCs mobilized in 1 apheresis day with M+G was 10.80x10 6 CD34+ cells/kg vs 2.14x10 6 CD34+ cells/kg with P+G. The # of cells infused was determined independently by each investigator according to local practice. Median time to neutrophil engraftment was 12 days in both arms (HR 0.94, 95% CI 0.62-1.41, p=0.75). Median time to platelet engraftment was 18 days (range: 17-19) with M+G and 17 days (range: 17-18) with P+G (HR 0.89, 95% CI 0.59-1.34, p=0.57). Graft durability at day 100 post-ASCT was 92.2% in the M+G arm and 91.9% in the P+G arm (OR 1.04, 95% CI 0.2-4.5, p=0.96). Overall, adverse events were reported in 98.8% (Grade 3/4: 68.8%) of patients with M+G vs 95.2% (Grade 3/4: 42.9%) with P+G, with cytopenias in the post-ASCT period prior to engraftment accounting for the majority of Grade 3/4 AEs in both arms, as expected. The most common AEs related to M included: local injection site reactions (any grade: 70.0%, Grade 3/4: 11.3%); and systemic reactions such as pruritis (33.8%), flushing (32.5%) and urticaria (12.5%). Additionally, mobilization with M+G resulted in a 10.5x increase in mean absolute # of CD34+CD45RA-CD123loCD38-CD90+CD49f+ primitive HSCs collected vs P+G (p<0.0001). Multicolor FACS and scRNA sequencing of CD34+ HSCs from both arms will be reported in a separate ASH abstract.
Conclusions: A single injection of M on top of G significantly increased the proportion of patients mobilizing ≥6x10 6 CD34+ cells/kg for ASCT (92.5%) vs G (26.2%) in up to 2 apheresis days (p<0.0001), while enabling 88.8% to collect ≥6x10 6 CD34+ cells/kg in just 1 apheresis (p<0.0001, Figure 1A). Despite the higher # of cells mobilized by M+G, the # of CD34+ cells/kg infused was determined independently by each investigator according to local practice with comparable engraftment kinetics and graft durability between the 2 arms. Finally, M+G mobilized 10.5x more immunophenotypically primitive CD34+ HSCs capable of durable multilineage hematopoietic engraftment vs P+G (p<0.0001, Figure 1B).
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Crees: BioLineRx Ltd.: Research Funding. Larson: TORL biotherapeutics: Current holder of individual stocks in a privately-held company; Bioline: Research Funding; Abbvie: Research Funding; BMS: Research Funding; Celgene: Research Funding; GSK: Research Funding; Janssen: Research Funding; Juno: Research Funding; Novartis: Research Funding; Pfizer: Research Funding; Takeda: Research Funding. Illés: Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees; Roche: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees. Stiff: Incyte: Research Funding; Cellectar: Research Funding; Seagen: Research Funding; Gamida Cell: Research Funding; Cellectar: Research Funding; Actinium: Research Funding; Bristol Myers Squibb: Research Funding; BioLineRX: Research Funding; Macrogenics: Research Funding; CRISPR Therapeutics: Consultancy, Honoraria; Amgen: Research Funding; Janssen: Research Funding; Kite, a Gilead Company: Research Funding; Karyopharm: Consultancy, Honoraria; MorphoSys: Consultancy, Honoraria. Sborov: Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees; GlaxoSmithKline: Consultancy; Sanofi: Consultancy; SkylineDx: Consultancy. Pereira: Jazz Pharmaceutical: Membership on an entity's Board of Directors or advisory committees. Mikala: Novartis: Consultancy; Takeda: Consultancy; Abbvie: Consultancy; Krka: Consultancy; Janssen: Consultancy; Amgen: Consultancy; Celgene: Consultancy. Holtick: Celgene: Honoraria; Sanofi: Honoraria. Qazilbash: Amgen: Research Funding; Oncopeptides: Other: Advisory Board; Bristol-Myers Squibb: Other: Advisory Board; Biolline: Research Funding; Angiocrine: Research Funding; NexImmune: Research Funding; Janssen: Research Funding. Hardy: American Gene Technologies, International: Membership on an entity's Board of Directors or advisory committees; Kite/Gilead: Membership on an entity's Board of Directors or advisory committees; InCyte: Membership on an entity's Board of Directors or advisory committees. Vainstein: BioLineRx LTD: Current Employment. Sorani: BioLineRx LTD: Current Employment. Gliko-Kabir: BioLineRx Ltd.: Current Employment. Goldstein: BioLineRx Ltd.: Current Employment. Kadosh: BioLineRx Ltd.: Current Employment.
Sodium and potassium-activated adenosine triphosphatase (Na
+, K
+-ATPase) and endogenous digitalis-like compounds (DLC) in the brain have been implicated in the pathogenesis of mood disorders. This ...hypothesis was examined by the determination of Na
+, K
+-ATPase/DLC system in parietal cortex of patients with different mood disorders and two animal models of depression.
Na
+, K
+-ATPase concentrations in human brain synaptosomal fractions, from patients with mood disorders, schizophrenia, and normal individuals, were determined by
3H-ouabain binding assay. Alpha isoforms were quantified by Western blotting. Brain DLC were measured using sensitive enzyme linked immunosorbant assay (ELISA). The effects of ouabain and ouabain-antibodies on behavior were determined in two animal models of depression.
3H-ouabain binding in bipolar patients was significantly lower than in major depressed and schizophrenic patients. Na
+, K
+-ATPase α isoforms in synaptosomal fractions were not different among the groups. DLC levels in the parietal cortex of bipolar patients were significantly higher than in normal individuals and depressed patients. Injection of lipopolysaccharide (intraperitoneally) to rats elicited depression-like symptoms, which were significantly attenuated by pre-injection of ouabain-antibodies. Injection of ouabain and ouabain-antibodies (intracerebroventricular) reduced depression-like symptoms in the forced swimming test in rats.
The results support the possibility that Na
+, K
+-ATPase and endogenous DLC participate in the pathogenesis of depressive disorders.
Abstract Depressive disorders are among the world's greatest public health problems. Na+ , K+ -ATPase is the established receptor for the steroidal digitalis-like compounds (DLC). Alteration in brain ...Na+ , K+ -ATPase and DLC have been detected in depressive disorders raising the hypothesis of their involvement in these pathology. The present study was designed to further elaborate this hypothesis by investigating the behavioral and biochemical consequences of neutralization in brain DLC activity attained by anti-ouabain antibodies administrations, in normal Sprague–Dawley (SD) and in the Flinders Sensitive Line (FSL) of genetically depressed rats. Chronic i.c.v. administration of anti-ouabain antibodies to FSL rats elicited anti-depressive behavior. Administration of anti-ouabain antibodies intracerebroventriculary (i.c.v.) to SD rats significantly changed the levels of catecholamines and their metabolites in the hippocampus, ventral tegmentum and nucleus accumbence. These results are in accordance with the notion that endogenous DLC may be involved in the manifestation of depressive disorders and suggests that alteration in their levels may be of significant therapeutic value.
Background The sodium- and potassium-activated adenosine triphosphatase (Na+ , K+ -ATPase) is a major plasma membrane transporter for sodium and potassium. We recently suggested that bipolar ...disorders (BD) may be associated with alterations in brain Na+ , K+ -ATPase. We further conjectured that the differences in Na+ , K+ -ATPase in BD patients could result partially from genetic variations in Na+ , K+ -ATPase α isoforms. Methods To test our hypothesis, we undertook a comprehensive study of 13 tagged single nucleotide polymorphisms (SNPs) across the three genes of the brain α isoforms of Na+ , K+ - ATPase ( ATP1A1 , ATP1A2 , and ATP1A3 , which encode the three α isoforms, α1, α2, and α3, respectively) identified using HapMap data and the Haploview algorithm. Altogether, 126 subjects diagnosed with BD from 118 families were genotyped (parents and affected siblings). Both individual SNPs and haplotypes were tested for association using family-based association tests as provided in the UNPHASED and PBAT set of programs. Results Significant nominal association with BD was observed for six single SNPs (α1: rs11805078; α2: rs2070704, rs1016732, rs2854248, and rs2295623; α3: rs919390) in the three genes of Na+ , K+ -ATPase α isoforms. Haplotype analysis of the α2 isoform ( ATP1A2 gene) showed a significant association with two loci haplotypes with BD (rs2295623: rs2070704; global p value = .0198, following a permutation test). Conclusions This study demonstrates for the first time that genetic variations in Na+ , K+ -ATPase are associated with BD, suggesting a role of this enzyme in the etiology of this disease.
עבודה זו תבקש לבחון את פעולותיהם של החשמונאים הראשונים, החל מעלייתו של מתתיה החשמונאי למנהיגות וכלה בתקופת מלכותו של אלכסנדר ינאי, בכל הנוגע למעשים של כפייה דתית ושל גיור אותם יישמו החשמונאים ואשר ...כֻּוונו נגד תושבים נוכריים בערים ההלניסטיות בארץ ישראל וכן נגד אי אלו עמים יושבי הארץ המחברת יוצאת מנקודת הנחה כי למעשים הקשורים בכפייה דתית ובגיור ליהדות יתכנו שני מקורות השראה: האחד, הלכות הדת היהודית וחוקי היהודים העוסקים ביחס אל הנוכרים היושבים בארץ ישראל והשני, פוליטיקה הלניסטית השואבת מן העולם היווני-הלניסטי אשר אותה אִ מצו החשמונאיםמתוך היותם של החשמונאים מנהיגים אשר קמו מקרב העם היהודי בגין התקוממות כלל יהודית נגד רדיפותיו, פקודותיו וגזרותיו של השלטון הסלווקי, קלה היא ההנחה כי קנאותם לטהרת הארץ מאלילות נוכרית ומפולחן נוכרי היא אשר הניעה אותם במעשיהם. ואכן הנחה זו הנה ברירת המחדל במחקר. אף כי ישנה הסכמה בנוגע לתהליכים של הלניזציה תרבותית וחברתית אשר התרחשו בקרב החשמונאים ככל שהתבססו אלה בשלטון, המגמה בכל הנוגע לחקר הפוליטיקה החשמונאית בכלל ופעולות הכפייה הדתית אותן ישמו בפרט הנה לייחס את אלה ליהדותם של החשמונאים ולקנאותם לחוקי התורה.ל אף מראית העין באשר ליהדותם של החשמונאים ולחרדתם לחוקי התורה ולטהרת הארץ הבאות לידי ביטוי בהתנהגותם כלפי הנוכרים יושבי הערים ההלניסטיות בארץ ישראל ובהתנהלותם אל מול אותם עמים אשר הוכפפו לכאורה תחת חוקי היהדות, ולמרות שהמקורות העיקריים מהם ניתן ללמוד אודות פעולות קנאות אלה, היינו שני ספרי המקבים וחיבוריו של יוסף בן מתתיהו, מנסים ככל יכולתם להעביר את המסר הלאומני-יהודי הקנאי בפעולות אלה של החשמונאים, משערת המחברת כי דווקא העולם ההלניסטי הן במרחב הסורי והן ביוון גופא, הוא הוא אשר השפיע על מדיניותם של החשמונאים כלפי הנוכרים עמם באו במגע.החשמונאים עלו לשררה ביהודה כאשר למן ראשית פעילותם כיוונה אותם השאיפה הפוליטית להדיח מן המנהיגות את הזרם היהודי המיוון, זה האחראי לשינוי הפולחני בבית המקדש בירושלים בשל שיתוף הפעולה שלו עם השלטון הסלווקי בפרט ועם העולם ההלניסטי בכלל, ולהעלות במקומו למנהיגות זרם יהודי מסורתי-שמרני אשר עליו הם נמנו ועבורו נלחמו. החלפת מנהיגות אחת באחרת תוך שינוי פולחן אחד באחר בבית המקדש נבעה, אמנם, ממניעים יהודיים קנאיים אולם מניעים אלה היו אך הכלי באמצעותו נוצחה והוחלפה כהונה גדולה אחת באחרת. שעה שהתבססו החשמונאים בשלטון ועם חלוף הזמן המשיכה הפוליטיקה לכוון את פעילותם תוך שמצאו ב'כלי' הקנאות לתורה שימוש יעיל ומועיל לצורך התבססותם בשלטון ואף הרחבתו.במידה והקנאות לתורה ולטהרת הארץ הנה רק אמצעי למדיניות ההתרחבות החשמונאית, המדיניות עצמה שואבת ממקורות אחרים, חוץ יהודאיים – מקורות הלניסטיים.הרחבת תחום שלטונם של החשמונאים תוך כדי סיפוחם והכנעתם של עמים שונים יושבי הארץ ואף של אזרחי הפולֵ ייס ההלניסטיות בתחומה נעשתה באופן אשר עשוי להזכיר את ההתארגנות הפדרטיבית של הליגות היווניות ההלניסטיות, התארגנות אשר ניתן לזהותה אף במרחב הסלווקי עצמו. לדעתה של המחברת, במדיניות ההתרחבות החשמונאית והסיפוחים אשר נלוו אליה ניתן לזהות מדיניות פדרטיבית או, לחילופין, מדיניות של מעין ליגה אשר נוסדה על ידי המנהיגים החשמונאים תוך שהיא שואבת את השראתה מהעולם ההלניסטי הסובבעבודה זו מוצאת את הקשר בין מדיניות הסיפוחים החשמונאית אשר מחקה את הפוליטיקה ההלניסטית ובעיקר, את ההתארגנויות הפדרטיביות שבו, לבין פעולות הכפייה הדתית והגיור אותן בִ צעו החשמונאים בדרכם להתרחבות שלטונית וטריטוריאלית. לדעתה של המחברת, השימוש בפעולות של כפייה דתית וגיור אִ פשר למנהיגים החשמונאים לבנות ליגה יהודית חזקה, אשר התרחבותה הטריטוריאלית נעשתה במקביל להתרחבותה החוקתית. התרחבות חוקתית זו היא השואבת מהפוליטיקה ההלניסטית מצד אחד תוך שמשתמשת היא בחוקי היהודים מצד שני, ובכך מציעה היא את חוקי היהודים כחוקה לכל דבר.כל שמבקשת עבודה זו להציע היא נקודת מבט מעט שונה על מדיניותם של החשמונאים, מדיניות אשר אינה ניתנת להפרדה מהמרחב הסלווקי בו הם פעלו ומהעולם ההלניסטי עמו הם באו במגע. המחברת רואה בתהליך המדיני אשר התרחש ביהודה עם עליית החשמונאים למנהיגות בד בבד להחלשותו של השלטון הסלווקי במרחב תהליך טבעי שאינו בהכרח יחודי או יוצא דופן אלא זהה בעקרונותיו לתהליכים שהתרחשו במקומות אחרים במרחב הסלווקי ובקרב עמים שונים בו. יוצא דופן הוא השימוש בו עשו מנהיגים אלה אשר קמו ביהודה בדת היהודית; ושימוש זה הוא שעומד למבחן במחקר זה.
The conversion funnel is a model describing the stages consumers go through in their journey toward a purchase. This journey often lasts several days to weeks and can include multiple visits to a ...seller’s website. A large body of literature has focused on using observable search patterns to identify consumers’ hidden purchasing stages and to estimate their likelihood of conversion. We propose a novel set of measures to better reveal the consumer’s hidden stage in the funnel. These measures are based on the diversity of the searches that a customer engages in while browsing an e-commerce website, and they include not only the number of different products that are searched for, but also measures that rely on unobserved similarities among products, captured in a product network (in which products are assumed to be “similar” if they are frequently co-searched). We operationalize and evaluate our proposed measures using a large-scale dataset from a medium-sized tourism website used for comparing and booking flights. We estimate a hidden Markov model to show that our proposed diversity measures are associated with progress in the funnel and consumers’ conversion likelihood. Specifically, we show that consumers go through different distinguishable stages (states) in their journey, characterized by different values of our proposed diversity measures. To demonstrate the managerial and business implications of our theory, we show that incorporating search-diversity measures into a baseline prediction model significantly improves the model’s performance in predicting purchase likelihood and churn.
Dysfunction of the human voltage‐gated K+ channel Kv1.1 has been associated with epilepsy, multiple sclerosis, episodic ataxia, myokymia, and cardiorespiratory dysregulation. We report here that ...AETX‐K, a sea anemone type I (SAK1) peptide toxin we isolated from a phage display library, blocks Kv1.1 with high affinity (Ki ~ 1.6 pM) and notable specificity, inhibiting other Kv channels we tested a million‐fold less well. Nuclear magnetic resonance (NMR) was employed both to determine the three‐dimensional structure of AETX‐K, showing it to employ a classic SAK1 scaffold while exhibiting a unique electrostatic potential surface, and to visualize AETX‐K bound to the Kv1.1 pore domain embedded in lipoprotein nanodiscs. Study of Kv1.1 in Xenopus oocytes with AETX‐K and point variants using electrophysiology demonstrated the blocking mechanism to employ a toxin‐channel configuration we have described before whereby AETX‐K Lys23, two positions away on the toxin interaction surface from the classical blocking residue, enters the pore deeply enough to interact with K+ ions traversing the pathway from the opposite side of the membrane. The mutant channel Kv1.1‐L296F is associated with pharmaco‐resistant multifocal epilepsy in infants because it significantly increases K+ currents by facilitating opening and slowing closure of the channels. Consistent with the therapeutic potential of AETX‐K for Kv1.1 gain‐of‐function‐associated diseases, AETX‐K at 4 pM decreased Kv1.1‐L296F currents to wild‐type levels; further, populations of heteromeric channels formed by co‐expression Kv1.1 and Kv1.2, as found in many neurons, showed a Ki of ~10 nM even though homomeric Kv1.2 channels were insensitive to the toxin (Ki > 2000 nM).
Kv1.1 channels are expressed in axons and dendrites of neurons. Increased Kv1.1 activity is associated with epilepsy. We found sea anemone toxin peptide AETX‐K blocks Kv1.1 with picomolar affinity and superb specificity, using its Lys23 to occlude the channel pore. We determined the AETX‐K structure and showed that 4 pM decreased the currents of Kv1.1‐L296F, a gain‐of‐function mutation associated with pharmaco‐resistant multifocal epilepsy, to wild‐type levels. AETX‐K blocks Kv1.1‐Kv1.2 heteromeric channels with a Ki of ~10 nM.
Cannabidiol-enriched oil (CBDO) is being used increasingly to improve seizure control in adult patients with drug-resistant epilepsy (DRE), despite the lack of large-scale studies supporting its ...efficacy in this patient population. We aimed to assess the effects of add-on CBDO on seizure frequency as well as on gait, cognitive, affective, and sleep-quality metrics, and to explore the electrophysiological changes in responder and non-responder DRE patients treated with add-on CBDO.
We prospectively recruited adult DRE patients who were treated with add-on CBDO. Patients were evaluated prior to treatment and following 4 weeks of a maintenance daily dose of ≈260 mg CBD and ≈12 mg Δ9-tetrahydrocannabinol (THC). The outcome measures included seizure response to CBDO (defined as ≥50% decrease in seizures compared to pre-CBDO baseline), gait testing, Montreal Cognitive Assessment (MoCA), Hospital Anxiety and Depression Scale (HADS), and sleep-quality questionnaire assessments. Patients underwent electroencephalography (EEG) recording during rest as well as event-related potentials (ERPs) during visual Go/NoGo task while sitting and while walking.
Nineteen patients were recruited, of which 16 finished pre- and post-CBDO assessments. Seven patients (43.75%) were responders demonstrating an average reduction of 82.4% in seizures, and nine patients (56.25%) were non-responders with an average seizure increase of 30.1%. No differences in demographics and clinical parameters were found between responders and non-responders at baseline. However, responders demonstrated better performance in the dual-task walking post-treatment (p = .015), and correlation between increase in MoCA and seizure reduction (r = .810, p = .027). Post-CBDO P300 amplitude was lower during No/Go-sitting in non-responders (p = .028) and during No/Go-walking in responders (p = .068).
CBDO treatment can reduce seizures in a subset of patients with DRE, but could aggravate seizure control in a minority of patients; yet we found no specific baseline clinical or electrophysiological characteristics that are associated with response to CBDO. However, changes in ERPs in response to treatment could be a promising direction to better identify patients who could benefit from CBDO treatment.
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