Citations

Purpose: The goal of this study was to develop a lot release assay for iPSC residuals following directed differentiation of iPSCs to retinal pigment epithelial (RPE) cells. Methods: RNA Sequencing (RNA Seq) of iPSCs and RPE derived from them was used to identify pluripotency markers downregulated in RPE cells. Quantitative real time PCR (qPCR) was then applied to assess iPSC residuals in iPSC-derived RPE. The limit of detection (LOD) of the assay was determined by performing spike-in assays with known quantities of iPSCs serially diluted into an RPE suspension. Results: ZSCAN10 and LIN28A were among 8 pluripotency markers identified by RNA Seq as downregulated in RPE. Based on copy number and expression of pseudogenes and lncRNAs ZSCAN10 and LIN28A were chosen for use in qPCR assays for residual iPSCs. Reverse transcription PCR indicated generally uniform expression of ZSCAN10 and LIN28A in 21 clones derived from 8 iPSC donors with no expression of either in RPE cells derived from 5 donor lines. Based on qPCR, ZSCAN10 , and LIN28A expression in iPSCs was generally uniform. The LOD for ZSCAN10 and LIN28A in qPCR assays was determined using spike in assays of RPE derived from 2 iPSC lines. Analysis of ΔΔC t found the limit of detection to be <0.01% of cells, equivalent to <1 iPSC/10,000 RPE cells in both iPSC lines. Conclusions: qPCR for ZSCAN10 and LIN28A detects <1 in 10,000 residual iPSCs in a population of iPSC-derived RPE providing an adequate LOD of iPSC residuals for lot release testing.

Abstract Background: Hispanic American (HA) men with prostate cancer (PC) are more likely to have more aggressive and advanced diseases and greater mortality rates compared to non-Hispanic white (NHW) men. Resistance to the current anticancer therapy is the major hurdle that leads to poor clinical outcomes. Hedgehog, Notch, and Wnt signaling pathways play a crucial role in development, progression, and resistance to the various anticancer therapies. Here, we characterized the enrichment of Hedgehog, Notch, and Wnt signaling pathways and molecular tumors differences between HA and NHW patients. Methods: Our data consist of 88 PC tumors samples (HA=34, NHW=54) obtained from treatment-naive metastatic hormone-sensitive prostate cancer (mHSPC) that were analyzed by next-generation sequencing (592, NextSeq; Whole Exome Sequencing, NovaSeq) and Whole Transcriptome Sequencing (WTS; NovaSeq) (Caris Life Sciences, Phoenix, AZ). Pathway enrichment was determined by Gene Set Enrichment Analysis (GSEA, Broad Institute) by measurable TPM (transcripts per kilobase million). The medical records were reviewed in a deidentified fashion for clinical features. Statistical significance was determined using chi-square and Mann-Whitney U (p&lt;0.05). Results: HA PC had enrichment of Hedgehog signaling (NES: 1.45, FDR=0.04), Notch signaling (NES: 1.41, FDR=0.07), and Wnt/β-catenin signaling (NES: 1.38, FDR=0.11) pathways compared to NHW tumors. HA tumors had higher expression of genes (HES1, Notch1, WNT1, WNT2; FC:1.1-2.5) associated with crosstalk between Hedgehog, Notch, and Wnt/β- catenin signaling pathways (all p&lt;0.05). Moreover, HA tumors had higher expression of stemness (ALDH1A2, ALDH1A1, PROM1; FC: 1.7-3.0), drug efflux (ABCC1, ABCB1; FC: 1.1- 1.5), epithelial to mesenchymal transition (EMT) (TWIST1, MMP2; FC: 1.1-1.5), cell cycle (CCND2; FC: 1.2), inhibition of apoptosis (BIRC2, XIAP; FC: 1.1) and epigenetic (EP300, TET1, TET2; FC: 1.3-1.4) related genes compared to NHW tumors (all p&lt;0.05). HA PC had higher frequency of Hedgehog and Wnt/β-catenin associated SPOP (16% vs 6.6%, p=0.2) and CTNNB1 (12% vs 2.2%, p=0.1) mutation compared to NHW PC. Conclusion: These results demonstrate that tumors from HA patients had enrichment of oncogenic Hedgehog, Notch, and Wnt/β-catenin signaling pathways, higher expression of stemness, drug efflux, EMT, cell cycle, inhibition of apoptosis and epigenetic-related genes, and mutations directly involved in activating the Hedgehog and Wnt/β-catenin signaling pathways. Together, these findings suggest that tumors from HA patients are enriched for gene signatures that promote cancer progression and therapy resistance. Additional research on collaboration and crosstalk between Hedgehog, Notch, and Wnt/β-catenin signaling pathways may help in designing better therapeutic strategies for HA men with mHSPC. Citation Format: Alejandro Recio-Boiles, Sachin Kumar Deshmukh, Ricardo J. Estrada-Mendizabal, Kenneth Barker, Sharon Wu, Joanne Xiu, Milan Radovich, Elisabeth I. Heath, Rana McKay, Juan Chipollini, Cynthia K. Miranti. Defining the role of Hedgehog, Notch, and Wnt signaling pathways in aggressive lethal prostate cancer of Hispanic men [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C100.

AbstractObjectivesAdolescents with functional abdominal pain (FAP) often experience pain and other gastrointestinal symptoms in the context of eating, which may place them at risk for eating disorders. This study compared disordered eating and its sequalae in adolescents with FAP to those with chronic headache.MethodsParticipants in this retrospective chart review study were 270 adolescents and young adults (mean age 15.9; standard deviation [SD] = 1.63; 60% female) attending an intensive, interdisciplinary pain treatment program, including 135 youth with FAP and an age‐ and gender‐matched control group with a primary pain diagnosis of chronic headache. Information was extracted related to eating‐ and weight‐related symptoms.ResultsIndependent t‐tests and χ2 analyses found that compared with adolescents with chronic headache, patients with FAP showed significantly more eating pathology as reflected by scores on the Eating Attitudes Test—26 (EAT‐26) (p < 0.001), endorsement of pain‐related restrictive eating (p < 0.01), eating disorder history (p < 0.001), and avoidant restrictive food intake disorder (ARFID) diagnoses (p < 0.001). Compared to those with chronic headache, significantly more patients with FAP had lost 10 pounds or more (p < 0.01) and a higher proportion were underweight (p < 0.001). Among all participants, a history of exclusion diets used to manage symptoms significantly increased the risk for ARFID (p < 0.001).ConclusionThe results of this study highlight the increased risk of eating‐related concerns among youth with FAP, as well as the judicious use of exclusion diets to manage symptoms in these patients.

Abstract Background Immune checkpoint inhibitors (ICIs) have transformed cancer treatment by targeting immune checkpoints such as PD‐1, PDL‐1, and CTLA‐4, but concerns about severe immune‐related adverse events persist. The scarcity of literature on dermatologic implications, especially severe reactions such as Stevens‐Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN), highlights the urgent need for investigation. Objective Our systematic review aims to address the gap in relevant literature by extensively examining the epidemiologic risk factors and management of SJS/TEN‐like illnesses in ICI‐treated patients to provide insights for risk assessment and clinical care. Method s We identified 158 case reports that detailed the incidence of SJS/TEN in patients being treated with ICIs, examining demographic patterns, type of malignancy, clinical characteristics, and treatments linked to onset. We assessed mortality rates, risk elements, and the effectiveness of interventions to help guide clinical care. Results Analysis of 158 case reports revealed that SJS/TEN in ICI users is typically seen on average at the age of 63 and is more common in males. PD1 inhibitors such as nivolumab and pembrolizumab are often associated with various mucocutaneous patterns and significant risks with ICI use, especially TEN, which is linked to high morbidity and mortality rates. Limitations Our study notes limitations due to the inclusion of case reports or case series, such as potential publication and reporting biases, leading to skewed findings. Additionally, because of the heterogeneous reporting standards, the retrospective nature limits phenotypic precision, control for confounding variables, and data completeness. Conclusion Our study provides valuable insights into the epidemiology, clinical features, management strategies, and outcomes of ICI‐induced SJS/TEN, underscoring the importance of vigilant monitoring and personalized risk assessment in oncology practice. Continued research efforts are essential to optimize patient outcomes and enhance the safety profile of ICIs in cancer therapy.

PURPOSE Imlunestrant is a next-generation oral selective estrogen receptor (ER) degrader designed to deliver continuous ER target inhibition, including in ESR1- mutant breast cancer. This phase Ia/b trial determined the recommended phase II dose (RP2D), safety, pharmacokinetics, and efficacy of imlunestrant, as monotherapy and in combination with targeted therapy, in ER-positive (ER+) advanced breast cancer (ABC) and endometrial endometrioid cancer. The ER+/human epidermal growth factor receptor 2–negative (HER2–) ABC experience is reported here. METHODS An i3+3 dose-escalation design was used, followed by dose expansions of imlunestrant as monotherapy or in combination with abemaciclib with or without aromatase inhibitor (AI), everolimus, or alpelisib. Imlunestrant was administered orally once daily and with the combination partner per label. RESULTS Overall, 262 patients with ER+/HER2– ABC were treated (phase Ia, n = 74; phase Ib, n = 188). Among patients who received imlunestrant monotherapy (n = 114), no dose-limiting toxicities or discontinuations occurred. At the RP2D (400 mg once daily), patients (n = 51) reported grade 1-2 nausea (39.2%), fatigue (39.2%), and diarrhea (29.4%). Patients at RP2D had received previous cyclin-dependent kinase 4/6 inhibitor (CDK4/6i; 92.2%), fulvestrant (41.2%), and chemotherapy (29.4%) for ABC and achieved a median progression-free survival (mPFS) of 7.2 months (95% CI, 3.7 to 8.3). Among patients who received imlunestrant + abemaciclib (n = 42) and imlunestrant + abemaciclib + AI (n = 43), most (69.4%) were treatment-naïve for ABC; all were CDK4/6i-naïve. Patients treated with imlunestrant + everolimus (n = 42)/alpelisib (n = 21) had received previous CDK4/6i (100%), fulvestrant (34.9%), and chemotherapy (17.5%) for ABC. No new safety signals or interactions with partnered drugs were observed. The mPFS was 19.2 months (95% CI, 13.8 to not available) for imlunestrant + abemaciclib and was not reached for imlunestrant + abemaciclib + AI. The mPFS with imlunestrant + everolimus/alpelisib was 15.9 months (95% CI, 11.3 to 19.1)/9.2 months (95% CI, 3.7 to 11.1). Antitumor activity was evident regardless of ESR1 mutation status. CONCLUSION Imlunestrant, as monotherapy or in combination with targeted therapy, had a manageable safety profile with evidence of preliminary antitumor activity in ER+/HER2– ABC.

AbstractBackgroundThe first commercially available photon‐counting‐detector CT (PCD‐CT) has been introduced for clinical use. However, its spectral performance on single‐ and dual‐contrast imaging tasks has not been comprehensively assessed.PurposeTo evaluate the spectral imaging performance of a clinical PCD‐CT system for single‐contrast material [iodine (I) or gadolinium (Gd)] and dual‐contrast materials (I and Gd) in comparison with a dual‐source dual‐energy CT (DS‐DECT).MethodsIodine (5, 10, and 15 mg/mL) and gadolinium (3.3, 6.6, and 9.9 mg/mL) samples, and their mixtures (I/Gd: 5/3.3 and 10/6.6 mg/mL) were prepared and placed in two torso‐shaped water phantoms (lateral dimensions: 30 and 40 cm). These phantoms were scanned on a PCD‐CT (NAEOTOM Alpha, Siemens) at 90, 120, and 140 kV. The same phantoms were scanned on a DS‐DECT (SOMATOM Force, Siemens) with 70/Sn150, 80/Sn150, 90/Sn150, and 100/Sn150 kV. The radiation dose levels were matched [volume CT dose index (CTDIvol): 10 mGy for the 30 cm phantom and 20 mGy for the 40 cm phantom] across all tube voltage settings and between scanners. Two‐material decomposition (I/water or Gd/water) was performed on iodine or gadolinium samples, and three‐material decomposition (I/Gd/water) on both individual samples and mixtures. On each decomposed image, mean mass concentration (± standard deviation) was measured in circular region‐of‐interests placed on the contrast samples. Root‐mean‐square‐error (RMSE) values of iodine and gadolinium concentrations were reported based on the measurements across all contrast samples and repeated on 10 consecutive slices.ResultsFor all material decomposition tasks on the DS‐DECT, the kV pairs with greater spectral separation (70/Sn150 kV and 80/Sn150 kV) yielded lower RMSE values than other DS‐DECT and PCD‐CT alternatives. Specifically, for the optimal 70/Sn150 kV, RMSE values were 1.2 ± 0.1 mg/mL (I) for I/water material decomposition, 1.0 ± 0.1 mg/mL (Gd) for Gd/water material decomposition, and 4.5 ± 0.2 mg/mL (I) and 3.7 ± 0.2 mg/mL (Gd), respectively, for I/Gd/water material decomposition. On the PCD‐CT, the optimal tube voltages were 120 or 140 kV for I/water decomposition with RMSE values of 2.0 ± 0.1 mg/mL (I). For Gd/water decomposition on PCD‐CT, the optimal tube voltage was 140 kV with gadolinium RMSE values of 1.5 ± 0.1 mg/mL (Gd), with the 90 kV setting on PCD‐CT generating higher RMSE values for gadolinium concentration compared to all DS‐DECT and PCD‐CT alternatives. For three material decomposition, both imaging modalities demonstrated substantially higher RMSE values for iodine and gadolinium, with 90 kV being the optimal tube potential for Gd/I quantitation on PCD‐CT [5.4 ± 0.3 mg/mL (I) and 3.9 ± 0.2 mg/mL (Gd)], and DS‐DECT at 100/Sn150 kV having larger RMSE values for both materials compared to the alternatives for either modality.ConclusionOptimal tube voltage for material decomposition on the clinical PCD‐CT is task‐dependent but inferior to DS‐DECT using 70/Sn150 kV or 80/Sn150 kV in two‐material decomposition for single‐contrast imaging (iodine/water or gadolinium/water). Three material decomposition (iodine/gadolinium/water) in dual‐contrast imaging yields substantially higher RMSE for both imaging platforms.

AbstractPurposeA dual‐source CT system can be operated in a high‐pitch helical mode to provide a temporal resolution of 66 ms, which reduces motion artifacts in CT pulmonary angiography (CTPA). It can also be operated in a multi‐energy (ME) mode to provide iodine maps, beneficial in the evaluation of pulmonary embolism (PE). No energy‐integrating detector (EID) CT can perform simultaneous ME and high‐pitch acquisition. This phantom study aimed to evaluate the ability of a photon‐counting‐detector (PCD) CT to perform simultaneous high‐pitch and ME imaging for CTPA.MethodsA motion phantom was used to mimic the respiratory motion. Two tubes filled with iodine with intravascular thrombus mimicked by injecting glue within the tubes were placed along with 5, 10, and 15 mg/mL iodine samples, on a motion phantom at 20 and 30 revolutions per minute. Separate high‐pitch and ME EID‐CT scans and a single high‐pitch ME PCD scan were acquired and virtual monoenergetic images and iodine maps reconstructed. Percent thrombus occlusion was measured and compared between static and moving images.ResultsWhen there was motion, EID‐CT ME suffered from significant motion artifacts. The measured iodine concentrations with PCD‐CT in high‐pitch ME were more stable when there was a motion, with a lower standard deviation than EID‐CT in ME mode. The estimated percent thrombus occlusion dropped significantly with applied motion on EID‐CT, while PCD‐CT high‐pitch ME mode showed good agreement between measurements on static or moving images.ConclusionPCD‐CT with combined ME and high‐pitch mode facilitates simultaneous accurate iodine quantification and assessment of intravascular occlusion.

Bi-allelic disruptive variants (nonsense, frameshift, and splicing variants) in KDM5B have been identified as causative for autosomal recessive intellectual developmental disorder type 65. In contrast, dominant variants, usually disruptive as well, have been more difficult to implicate in a specific phenotype, since some of them have been found in unaffected controls or relatives. Here, we describe individuals with likely pathogenic variants in KDM5B, including eight individuals with dominant missense variants. This study is a retrospective case series of 21 individuals with variants in KDM5B. We performed deep phenotyping and collected the clinical information and molecular data of these individuals’ family members. We compared the phenotypes according to variant type and to those previously described in the literature. The most common features were developmental delay, impaired intellectual development, behavioral problems, autistic behaviors, sleep disorders, facial dysmorphism, and overgrowth. DD, ASD behaviors, and sleep disorders were more common in individuals with dominant disruptive KDM5B variants, while individuals with dominant missense variants presented more frequently with renal and skin anomalies. This study extends our understanding of the KDM5B-related neurodevelopmental disorder and suggests the pathogenicity of certain dominant KDM5B missense variants.

BACKGROUND: Whether vigorous exercise increases risk of ventricular arrhythmias for individuals diagnosed and treated for congenital long QT syndrome (LQTS) remains unknown. METHODS: The National Institutes of Health–funded LIVE-LQTS study (Lifestyle and Exercise in the Long QT Syndrome) prospectively enrolled individuals 8 to 60 years of age with phenotypic and/or genotypic LQTS from 37 sites in 5 countries from May 2015 to February 2019. Participants (or parents) answered physical activity and clinical events surveys every 6 months for 3 years with follow-up completed in February 2022. Vigorous exercise was defined as ≥6 metabolic equivalents for >60 hours per year. A blinded Clinical Events Committee adjudicated the composite end point of sudden death, sudden cardiac arrest, ventricular arrhythmia treated by an implantable cardioverter defibrillator, and likely arrhythmic syncope. A National Death Index search ascertained vital status for those with incomplete follow-up. A noninferiority hypothesis (boundary of 1.5) between vigorous exercisers and others was tested with multivariable Cox regression analysis. RESULTS: Among the 1413 participants (13% <18 years of age, 35% 18–25 years of age, 67% female, 25% with implantable cardioverter defibrillators, 90% genotype positive, 49% with LQT1, 91% were treated with beta-blockers, left cardiac sympathetic denervation, and/or implantable cardioverter defibrillator), 52% participated in vigorous exercise (55% of these competitively). Thirty-seven individuals experienced the composite end point (including one sudden cardiac arrest and one sudden death in the nonvigorous group, one sudden cardiac arrest in the vigorous group) with overall event rates at 3 years of 2.6% in the vigorous and 2.7% in the nonvigorous exercise groups. The unadjusted hazard ratio for experience of events for the vigorous group compared with the nonvigorous group was 0.97 (90% CI, 0.57–1.67), with an adjusted hazard ratio of 1.17 (90% CI, 0.67–2.04). The upper 95% one-sided confidence level extended beyond the 1.5 boundary. Neither vigorous or nonvigorous exercise was found to be superior in any group or subgroup. CONCLUSIONS: Among individuals diagnosed with phenotypic and/or genotypic LQTS who were risk assessed and treated in experienced centers, LQTS-associated cardiac event rates were low and similar between those exercising vigorously and those not exercising vigorously. Consistent with the low event rate, CIs are wide, and noninferiority was not demonstrated. These data further inform shared decision-making discussions between patient and physician about exercise and competitive sports participation. REGISTRATION: URL: https://www.clinicaltrials.gov ; Unique identifier: NCT02549664.

Abstract Objectives To examine the clinicopathologic features of patients with polymyalgia rheumatica (PMR) who had thoracic aorta repair surgery. Findings were compared with those of a cohort of patients with giant cell arteritis (GCA) requiring thoracic aorta repair. Methods All patients evaluated at Mayo Clinic in Rochester, MN, with Current Procedural Terminology (CPT) codes for thoracic aorta repair surgery between 2000–2021 were identified. All patients were screened for prior PMR diagnosis. Patients with PMR and no signs of GCA were categorized as clinically isolated PMR. The medical records of all patients were manually reviewed, and pathologists re-examined all the aortic tissues. Results Of the 4621 patients with at least one CPT code for thoracic aorta repair surgery, 43 patients were diagnosed with clinically isolated PMR before the surgery. Detailed histopathological examination of the aortic tissues revealed active inflammation in 30/43 (70%) patients after a median (IQR) of 10.0 (4.7–13.3) years from the PMR diagnosis. When compared with aortic tissue from patients with a prior diagnosis of GCA, the aorta of patients with PMR had more severe inflammation (Grade 3: 15/30 [50%] vs 5/34 [15%], P = 0.002). Patients with PMR and thoracic aorta repair may experience a 40% increased risk of mortality compared with the general population, but this did not reach statistical significance (standardized mortality ratio: 1.40; 95% CI: 0.91–2.07). Conclusions Some patients with PMR have subclinical aortic inflammation that is detectable many years after initial diagnosis and may contribute to the development of aortic aneurysm.