Multiple Lots Research Articles

7953 Testosterone II (TSTII) Immunoassay Meets the CDC HoSt Certification Requirements for a Fourth Consecutive Year

Abstract Disclosure: K.N. Nissen: Employee; Self; Siemens Healthineers. N. Parker: Employee; Self; Siemens Healthineers. P. Samantha: Employee; Self; Siemens Healthineers. J.M. Rhea-McManus: Employee; Self; Siemens Healthineers. V. Shalhoub: Employee; Self; Siemens Healthineers. J. Freeman: Employee; Self; Siemens Healthineers. Background: The Atellica® IM and ADVIA Centaur® Testosterone II (TSTII) assay is enrolled in the CDC Hormone Standardization (CDC HoST) Certification Program, which is traceable to the primary testosterone Standard National Measurement Institute (NMI) M914. Certification is a yearlong process of 40 blinded samples from the CDC, 10 to be tested each quarter. If 4 consecutive quarters meet the bias acceptance criteria of ± 6.4%, CDC certification is achieved. The TSTII assay is standardized using internal standards made from United States Pharmacopea (USP)-grade testosterone. Standards are traceable to ID-LC-MS/MS by method comparison using CDC samples; ID-LC-MS/MS is traceable to the primary testosterone standard NMI M914. Methods: Initially, to standardize the assay, approximately 120 samples were obtained from CDC with CDC HoST ID-LC-MS dose values to value assign master curve standards. Multiple ADVIA Centaur TSTII reagent lots were tested with the CDC patient samples. Internal master curve standards were assayed as unknowns and dose values assigned using the RLU (system generated units–Relative Light Units) to CDC ID-LC-MS dose relationship. From 2019 to 2023, after standard value assignment, at least 179 individual patient samples from the CDC with blinded amounts of testosterone have been tested across all platforms. Each quarter, 10 blinded samples were tested in duplicate over 2 days. Results were analyzed by the CDC according to CLSI EP9-A2 for bias. Results: Correlation between the CDC assigned values and the Atellica IM, ADVIA Centaur, and ADVIA Centaur CP TSTII assay results was strong across all quarterly blind trials from 2019 to 2023 combined—Atellica IM intercept = +0.5 ng/dL, slope = 1.03, r = 0.990, n = 180, Bland Altman mean bias was 4.55%; ADVIA Centaur intercept = +0.3 ng/dL, slope = 1.04, r = 0.993, n = 179, mean bias was -0.27%; ADVIA Centaur CP intercept = +0.9 ng/dL, slope = 1.07, r = 0.993, n = 180, mean bias was 1.26%. The TSTII assay bias on all systems was consistently <6.4%. This enabled the TSTII assays to be CDC certified for a fourth consecutive year. Conclusions: The TSTII assay has consistently achieved CDC certification from 2019 to 2023. Siemens Healthineers is the only testosterone immunoassay manufacturer to be currently certified and has achieved certification for four consecutive years. Presentation: 6/1/2024

A-065 The CDC HoSt Certification Requirements Have Been Met for More Than Four Years by the Atellica IM, ADVIA Centaur Testosterone II and Dimension Vista LOCI Total Testosterone Immunoassays

Abstract Background The CDC Hormone Standardization (CDC HoST) Certification Program, is traceable to the primary testosterone Standard National Measurement Institute (NMI) M914. The Atellica® IM, ADVIA Centaur® Testosterone II (TSTII) and Dimension Vista® LOCI Total Testosterone (TTST) assays are enrolled in the program. Certification is a yearlong process of 40 blinded samples from the CDC, 10 for testing each quarter. If 4 consecutive quarters meet bias acceptance criteria of ± 6.4%, CDC certification is achieved. Methods Initially, to standardize the assay approximately 120 samples were obtained from CDC with CDC HoST ID-LC-MS dose values to value assign master curve standards. Multiple ADVIA Centaur TSTII reagent lots were tested with CDC patient samples. Internal master curve standards were assayed as unknowns and dose values assigned using the RLU (system generated units-Relative Light Units) to CDC ID-LC-MS dose relationship. For TSTII (2019- 2023), after standard value assignment, at least 179 CDC blinded samples were tested across all platforms. Each quarter, 10 blinded samples were tested in duplicate over 2 days. Similar procedures were followed for TTST. Results were analyzed by the CDC. Results Correlation between the CDC assigned values and Atellica IM, ADVIA Centaur, and ADVIA Centaur CP TSTII assay results was strong across all quarters from 2019 to 2023 combined. TTST results were strong for 2023. Mean bias between TSTII and TTST results and CDC reference values was consistently <6.4%. This enabled the TSTII assays to be CDC certified for a fourth consecutive year, and TTST for 2023. The table presents TSTII (2019-2023) and TTST (2014-2023) results. Conclusions The TSTII assay consistently achieved CDC certification from 2019 to 2023, while the TTST assay was initially certified in 2014. Siemens Healthineers is the only testosterone immunoassay manufacturer to be currently certified, the TSTII assay has achieved certification for four consecutive years.

Analytical validation of the LungLB test: a 4-color fluorescence in-situ hybridization assay for the evaluation of indeterminate pulmonary nodules

BackgroundEvaluation of indeterminate pulmonary nodules (IPNs) often creates a diagnostic conundrum which may delay the early detection of lung cancer. Rare circulating genetically abnormal cells (CGAC) have previously demonstrated utility as a biomarker for discriminating benign from malignant small IPNs in the LungLB assay. CGAC are identified using a unique 4-color fluorescence in-situ hybridization (FISH) assay and are thought to reflect early cell-based events in lung cancer pathogenesis and the anti-tumor immune response. LungLB is a prognostic tool that combines the CGAC biomarker and clinical features to aid in IPN evaluation by improving the stratification of patient risk of malignancy.MethodsHerein we describe the analytical performance of the LungLB blood test. Analytical validation was performed according to Clinical and Laboratory Standards Institute (CLSI) guidelines with adaptations for rare cell-based assays. Multiple operators, reagent lots, and assay runs were tested to examine accuracy, precision, reproducibility, and interfering factors.ResultsThe FISH probes used in the LungLB assay demonstrate 100% sensitivity and specificity for their intended chromosomal loci (3q29, 3p22.1, 10q22.3 and 10cen). LungLB demonstrates analytical sensitivity of 10 CGAC per 10,000 lymphocytes analyzed, 100% analytical specificity, and high linearity (R2 = 0.9971). Within run measurements across 100 samples demonstrated 96% reproducibility. Interfering factors normally found in blood (lipemia, biotin) and exposure to adverse temperatures (-20ºC or 37ºC) did not interfere with results. Sample stability was validated to 96 hours.ConclusionThe analytical performance of LungLB in this validation study successfully demonstrates it is robust and suitable for everyday clinical use.

Analysis of therapeutic monoclonal antibodies by imaged capillary isoelectric focusing (icIEF).

Imaged capillary isoelectric focusing (icIEF) is a preferred analytical method for determining isoelectric points (pIs) and charge heterogeneity profiles in biotherapeutic proteins. In this study, we optimized the icIEF method for an in-house IgG1κ monoclonal antibody (mAb-1) and assessed its reproducibility, robustness, and autosampler stability. The optimized method was used to evaluate batch-to-batch consistency in pIs for multiple lots of mAb-1 and determine the relative percentages of charge variants. We also tested the method's performance using multiple lots of another IgG1 mAb, commercially available as Herceptin (trastuzumab). Additionally, we designed and assessed native and denaturing platform icIEF methods for 11 other marketed mAbs, with pIs ranging from 6.0 (eculizumab) to 9.22 (tocilizumab).

Dynamic Spatio-Temporal Adaptive Graph Convolutional Recurrent Networks for Vacant Parking Space Prediction

The prediction of vacant parking spaces (VPSs) can reduce the time drivers spend searching for parking, thus alleviating traffic congestion. However, previous studies have mostly focused on modeling the temporal features of VPSs using historical data, neglecting the complex and extensive spatial characteristics of different parking lots within the transportation network. This is mainly due to the lack of direct physical connections between parking lots, making it challenging to quantify the spatio-temporal features among them. To address this issue, we propose a dynamic spatio-temporal adaptive graph convolutional recursive network (DSTAGCRN) for VPS prediction. Specifically, DSTAGCRN divides VPS data into seasonal and periodic trend components and combines daily and weekly information with node embeddings using the dynamic parameter-learning module (DPLM) to generate dynamic graphs. Then, by integrating gated recurrent units (GRUs) with the parameter-learning graph convolutional recursive module (PLGCRM) of DPLM, we infer the spatio-temporal dependencies for each time step. Furthermore, we introduce a multihead attention mechanism to effectively capture and fuse the spatio-temporal dependencies and dynamic changes in the VPS data, thereby enhancing the prediction performance. Finally, we evaluate the proposed DSTAGCRN on three real parking datasets. Extensive experiments and analyses demonstrate that the DSTAGCRN model proposed in this study not only improves the prediction accuracy but can also better extract the dynamic spatio-temporal characteristics of available parking space data in multiple parking lots.

Incidence of any infections and SARS-CoV-2 infections in patients receiving treatment for follicular lymphoma and diffuse large B-cell lymphoma: Real-world evidence from a large US national claims database.

e19061 Background: Non-Hodgkin’s lymphoma is a heterogeneous group of conditions, including the indolent subtype follicular lymphoma (FL) and the aggressive subtype diffuse large B-cell lymphoma (DLBCL). Infections associated with NHL are a major concern due to effects of the underlying disease and its treatment and have been reported to increase with increasing lines of therapy (LoT). However, the risk of infections overall and SARS-COV-2 infections specifically is not well characterized in patients who have received multiple LoTs. The goal of the study was to estimate the exposure-adjusted incidence rates (EAIR) of infections during the COVID-19 pandemic by type, severity and LoT received in patients treated for FL or DLBCL in the real-world setting. Methods: We identified patients in the Optum Clinformatics claims data from October 1, 2015, to June 30, 2023. Patients were aged ≥18 years, enrolled continuously for 365 days before index (start of LoT), and had ≥2 ICD-10 diagnosis codes for FL or DLBCL in the 365 days before index. The EAIR (per 100 person-years, PY) was estimated for infections by type (any infections and SARS-COV-2 [after 1/1/2020]), severity (hospitalized or fatal hospitalizations), and number of LoTs received. Results: A total of 4,799, 1,025 and 510 FL patients were included in the 1L (1 LoT), 2L, and 3L+ cohorts, respectively. The EAIR of infections resulting in hospitalization in 1L, 2L, and 3L+ cohorts were 20.6 (95% confidence interval (CI): 19.0-22.3), 30.2 (25.5-35.9), and 44.2 (35.4-55.3) per 100-PY for any infection; and 4.6 (4.0-5.4), 4.4 (3.0-6.5) and 8.3 (5.3-13.2) per 100-PY for SARS-COV-2 infections, respectively. The EAIR of fatal hospitalizations were 4.0 (3.4-4.8), 8.6 (6.4-11.5), and 13.9 (9.7-20.0) per 100-PY for any infection; and 1.3 (1.0-1.7), 1.6 (0.8-3.0), and 4.1 (2.2-7.9) per 100-PY for SARS-COV-2 infections. A total of 8,058, 2,114 and 977 DLBCL patients were included in 1L, 2L, and 3L+ cohorts, respectively. The EAIR of infections resulting in hospitalization in 1L, 2L, and 3L+ cohorts were 64.2 (60.7-67.8), 85.2 (76.9-94.3), and 113.4 (98.4-130.7) per 100-PY for any infection; and 6.3 (5.5-7.2), 7.3 (5.6-9.6), and 15.1 (11.3-20.2) per 100-PY for SARS-COV-2 infections respectively. The EAIR of fatal hospitalizations were 12.0 (10.8-13.3), 23.0 (19.5-27.1), and 27.8 (22.1-34.9) per 100-PY for any infection; and 1.6 (1.2-2.1), 2.7 (1.7-4.2), and 4.8 (2.9-8.0) per 100-PY for SARS-COV-2 infections in 1L, 2L and 3L+ cohorts, respectively. Conclusions: This real-world analysis demonstrated substantial morbidity and mortality associated with overall and SARS-COV-2 infections among NHL patients, with a higher incidence in DLBCL compared to FL, and with increasing LoTs.

Map-based localization for intelligent vehicles based on fusion of multiple visual features in underground parking

The ‘last mile’ problem in intelligent car localization within underground parking lots presents challenges such as localization blind spots and feature extraction difficulties. To tackle these issues, this paper proposes a mapping and multiscale matching localization method based on the fusion of multiple feature sources using a ‘binocular mapping, monocular localization’ approach. By combining traditional features with deep learning features, the proposed method enhances feature matching accuracy and addresses feature extraction challenges. A sparse three-dimensional reconstruction method based on local feature-optical flow is also proposed to mitigate localization blind spots. During the mapping process, the underground parking lot scene is segmented into several nodes to store map information, each consisting of a feature layer, scene structure layer, and trajectory layer. In the multiscale matching localization process, ORB, LoFTR, and BoW multisource features are fused to achieve feature matching, determine the nearest node, and estimate vehicle pose using the PnP model through optical flow. Finally, a localization optimization algorithm outputs the localization result. Experiments were conducted across multiple underground parking lots encompassing a total area of over 100,000 square meters. The results demonstrated that the average error of map construction using the proposed algorithm is approximately 15 mm, and the average error rate of localization is 4.45%.

Abstract 4628: Validation of an automated, scalable comprehensive genomic profiling assay for hematologic malignancies

Abstract Optimal diagnosis, prognosis, and treatment selection for hematologic malignancies requires assessment of somatic mutations across a subset of clinically relevant genes. Here we present the analytical validation results of an expanded targeted next generation sequencing (NGS) panel of 141 clinically relevant genes for myeloid and lymphoid malignancies. Based on review of clinical guidelines, 141 genes including 79 myeloid and 84 lymphoid associated genes were selected. The expanded hematological panel interrogates all coding exons of the 141 genes to detect single nucleotide variants (SNVs) and insertions/deletions (indels). The panel also identifies copy number aberrations (CNAs) in 16 genes, FLT3 internal tandem duplicates (ITDs), select non-coding pathogenic variants, and patient sex. The custom hybrid capture-based assay utilizes genomic libraries created from 250 ng gDNA extracted from peripheral blood, bone marrow, or cell suspensions, followed by sequencing on Illumina® instruments. Concordance studies were performed on clinical samples previously assessed using an orthogonal NGS-based laboratory developed test for SNVs/indels/FLT3-ITDs or digital multiplexed ligation-dependent probe amplification for CNAs. In total, 308 patient samples were evaluated and included 72 (23.4%) acute myeloid leukemia and 58 (18.8%) myelodysplastic syndromes samples among other indications, of which 240 (77.9%) were bone marrow specimens. Analysis of concordance demonstrated a positive percent agreement (PPA) of 99.7% for SNVs (750/752), 99.5% for indels <25bp (206/207), and 95.7% for indels ≥ 25bp (22/23). Variants were from 45 genes including TP53, NPM1, and IDH1/2. PPA was 100% for FLT3-ITDs (38/38; size range 18-300 bp) and sex (294/294). For CNAs, PPA was 96.4% (132/137) and negative percent agreement (NPA) was 98.3% (931/947). Assay precision was determined using three replicates of 15 clinical samples for both intra and inter-assay precision using multiple operators, instruments, and reagent lots. 100% concordance was observed for SNVs (36/36), indels (11/11), a 24 bp FLT3-ITD (1/1), CNAs (24/24), and sex (15/15). Analytical specificity was assessed using 5 replicates of NA12878 showing specificity >99.99% for SNVs/indels with variant allele frequency ≥ 3%, and >99.99% for CNAs with copy number ≤0.85 or ≥1.15. Dilution series to determine analytical sensitivity are in progress. This study highlights the analytical validation of the expanded NGS panel for the detection of clinically informative genomic alterations in hematologic malignancies. Results from this validation study, when complete, will include at least 576 samples plus orthogonal testing. These data describe the performance of the assay to enable a comprehensive evaluation of genomic alterations using a single sample, further facilitating the use of broad NGS assays in patients with hematologic malignancies. Citation Format: Grant Hogg, Tong Liu, Helen Cao, Adib Shafi, Ashraf Shabaneh, Jon Williams, John Howitt, Amanda Williamson, Rachel Dango, Xiaojun Guan, Heidi Hoffmann, Michael Mooney, John Pruitt, Scott Parker, Henry Dong, Stan Letovsky, Li Cai, Eric A. Severson, Shakti H. Ramkissoon, Anjen Chenn, Marcia Eisenberg, Brian Caveney, Eyad Almasri, Taylor J. Jensen. Validation of an automated, scalable comprehensive genomic profiling assay for hematologic malignancies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4628.

Brassica Pharma recalls multiple lots of four eye ointments

Brassica Pharma recalls multiple lots of four eye ointments

Testing the test strips: laboratory performance of fentanyl test strips

BackgroundThe overdose crisis driven by synthetic opioids continues to escalate in the USA. We evaluated the efficacy of multiple manufacturing lots of a fentanyl test strip (FTS) to detect fentanyl and fentanyl analogs and assessed cross-reactivity with possible interferences.MethodsDrug standards were dissolved in water in a laboratory setting and serially diluted. Drug dilutions were tested using five different manufacturing lots of BTNX Rapid Response (20 ng/mL cutoff) lateral flow chromatographic immunoassay strips to assess lot-to-lot variability for FTS sensitivity and cross-reactivity for the analytes of interest.ResultsAll five manufacturing lots cross-reacted with fentanyl and eleven fentanyl analogs. Diphenhydramine, lidocaine, MDMA, and methamphetamine were found to cause false positives with the strips. There was notable lot-to-lot variability in the sensitivity of the strips for fentanyl, fentanyl analogs, and known interferences.DiscussionFTS remains an important overdose prevention tool, but lot-to-lot variability in performance complicates robust instructions that balance the prevention of false positives and false negatives. Continued lot-to-lot performance assessment is recommended to ensure health education for FTS remains accurate. More sophisticated drug checking technologies and services are needed in the community landscape to augment personal FTS use to facilitate informed consumption and overdose risk mitigation.

Realising transitions for urban regeneration through statutory planning: the case of greyfield precinct renewal in suburban Melbourne

ABSTRACT The compact city agenda aims to deliver sustainable future urban form. However, as with all planning, there is contestation. While built form and housing have primacy, cities need space for hard and soft infrastructural systems, including natural systems that, until recently, have been taken for granted but are diminishing through poorly planned densification. Nowhere is this more evident than in the suburbs, where lot-by-lot redevelopment is significantly reducing open space and tree canopy. Infill developments are not producing the benefits expected from compact cities; they are increasing strain on the grey, blue, and green infrastructure. This paper highlights limited intent in local planning regulations to promote anything other than business as usual, which, in the Australian context, is single lots becoming 2–4 subdivisions. While planning schemes can regulate negative externalities, they do not consider this when taken as a whole, across a redeveloped neighbourhood predominantly consisting of housing, and are not using the redevelopment opportunities to build a more resilient future for local areas. This paper builds on the Australian greyfield literature by showing how statutory amendments can promote precinct scale development, working across multiple lots to deliver housing concurrently with greater levels of amenity for existing and future residents. KEY POINTS Infill housing development on single lots is reducing local amenity while not making the fullest use of the land for housing supply. Site assembly precincts are critical for higher yield, higher amenity, urban redevelopment. Strategic policy alone will not deliver the solutions; statutory mechanisms for doing so are critical. This paper presents a fully worked statutory response to the issue, highlighting the method for developing the scheme and the future work necessary for delivering site-assembly precincts at council level.

Predicting Vacant Parking Space Availability Zone-Wisely: A Graph Based Spatio-Temporal Prediction Approach

Intelligent parking guidance systems (IPGSs) based on vacant parking space (VPS) availability predictions are highly effective to alleviate the increasingly serious parking difficulties in metropolis. The spatial-temporal correlation analysis of VPS information of multiple parking lots in a region shows that not only the number of VPS in each parking lot has a stable temporal correlation, but also there is obvious spatial correlation among different parking lots. Given this, this paper takes full advantage of the space-time correlation and makes short-term (within 30 min) and long-term (over 30 min) predictions about the number of VPSs in multiple parking lots in an area parallelly. Specifically, this paper develops a deep gated graph recurrent neural network (G <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex-math notation="LaTeX">$^{2}$</tex-math></inline-formula> RNN) model which has the ability to extract both spatial and temporal correlations concurrently between different parking lots. The advantages of the G <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex-math notation="LaTeX">$^{2}$</tex-math></inline-formula> RNN model are that, first, spatial and temporal correlations between different parking lots can be accurately obtained simultaneously, and second, there is no need to grid the raw data since it uses graph-structured data. For long-term predictions, two different approaches, namely the single-step and iterative predictions, are investigated. The performance of the G <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex-math notation="LaTeX">$^{2}$</tex-math></inline-formula> RNN based prediction method is extensively evaluated with practical data collected from eight public parking lots in Santa Monica. The results show that it can achieve considerably high accuracy in both short-term and long-term predictions.

Characterization and optimization of variability in a human colonic epithelium culture model.

Animal models have historically been poor preclinical predictors of gastrointestinal (GI) directed therapeutic efficacy and drug-induced GI toxicity. Human stem and primary cell-derived culture systems are a major focus of efforts to create biologically relevant models that enhance preclinical predictive value of intestinal efficacy and toxicity. The inherent variability in stem cell-based cultures makes development of useful models a challenge; the stochastic nature of stem cell differentiation interferes with the ability to build and validate reproducible assays that query drug responses and pharmacokinetics. In this study, we aimed to characterize and reduce sources of variability in a complex stem cell-derived intestinal epithelium model, termed RepliGut® Planar, across cells from multiple human donors, cell lots, and passage numbers. Assessment criteria included barrier for­mation and integrity, gene expression, and cytokine responses. Gene expression and culture metric analyses revealed that controlling cell passage number reduces variability and maximizes physi­ological relevance of the model. In a case study where passage number was optimized, distinct cytokine responses were observed among four human donors, indicating that biological variability can be detected in cell cultures originating from diverse human sources. These findings highlight key considerations for designing assays that can be applied to additional primary cell-derived systems, as well as establish utility of the RepliGut® Planar platform for robust development of human-pre­dictive drug-response assays.

대지사용권의 분유(分有)와 부당이득의 관계

According to the Supreme Court's unanimous decision on August 25, 2022(2017 DA 257067), the general legal principles regarding shared property in civil law cannot be directly applied to the “shared relationship” of collective building land. Therefore, co-owners of divided land, who hold a reasonable share corresponding to the ratio of exclusive parts in the collective building, have a legal authority to use and profit from the entire land according to its intended purpose. As a result, a land co-owner who is not a divided land owner cannot claim unjust enrichment against a divided land owner with a reasonable share based on the land co-owner's share rights. The conclusion of this unanimous decision by the Supreme Court emphasizes the obligation of unjust enrichment for divided landowners based on two factors: (1) the emphasis on the substantive use relationship of land use rights and (2) the economic viability of this conclusion. This perspective appears reasonable.&#x0D; The Supreme Court’s viewpoint, as presented in the unanimous decision, assumes the “shared relationship,” which is the typical ownership structure of land use rights in collective buildings. However, it is necessary to examine whether there is a change in legal relationships in cases where non-co-owners divide and separately (or jointly) own rights such as ownership of part of the land, apart from the shared relationship.&#x0D; The focus of this paper, the Supreme Court’s decision on November 28, 2019(2017 DA 294608), addresses issues related to unjust enrichment concerning land use rights when a single collective building is situated on multiple separate lots. While acknowledging the legal and economic validity of the unanimous decision, this paper critically analyzes the decision on the obligation of unjust enrichment. It suggests the need for legislative measures to clarify and advance future judgements or legal relationships regarding unjust enrichment in land-use relationships.

Patterns of Care and Resource Use Among Elderly Relapsed/Refractory Follicular Lymphoma Patients: US Medicare Claims Analysis

Background: Follicular lymphoma (FL)is the second most common type of non-Hodgkin lymphoma (NHL) and most common indolent NHL. Rituximab-based chemotherapy is highly effective and considered the mainstay first-line (1L) treatment for FL, though many patients relapse and continue through successive lines of therapy. These relapsed/refractory (R/R) patients experience worse outcomes and represent an unmet need for more effective treatment approaches. This study examined treatment patterns, health care resource utilization (HCRU), and costs among R/R FL patients initiating third line (3L) therapy between 2011 and 2020. Methods: This retrospective, observational study used Center for Medicare and Medicaid fee-for-service Medicare data. The sample included treated patients with FL (identified via 1 inpatient or 2 outpatient claims) aged ≥66 years who initiated 1, 2, and 3L therapy between January 1, 2011, and December 31, 2020. Patients were required to have continuous coverage in the 12 months pre-FL diagnosis and 3 months post-diagnosis and were followed until discontinuation of enrollment or death. Line of therapy (LoT) was calculated until the end of treatment, death, or transformation to diffuse large B-cell lymphoma. Outcomes included patient characteristics, treatment patterns, health care costs and HCRU. R esults: This analysis identified 8737 1L, 1847 2L, and 410 3L treated FL patients. Median age at the start of each LoT was 75 years at 1L, 77 at 2L, and 79 at 3L. Of R/R FL patients initiating 3L, the most common baseline Charlson comorbidities included diabetes without chronic complications (23%), chronic pulmonary disease (22%) and renal disease (20%), 32% were defined as having progression of disease within 2 years of 1L (POD24), and 30% were defined as double refractory to both an anti-CD20 agent and an alkylating agent. Treatment utilization rates were similar across LoTs; the most used regimens were rituximab/obinutuzumab (R/O) (used 35% of the time in 1L, 33% in 2L, and 31% in 3L) and bendamustine + R/O (39% in 1L, 33% in 2L, and 21% in 3L). Mean duration of 1L treatment was longer for patients who only initiated 1L (10.1 months) vs. those with R/R FL who went on to initiate 2L and 3L (7.5 and 7.4 months, respectively). As R/R FL patients who initiated 3L moved through LoTs, increases were observed in the number of patients with at least 1 inpatient admission (17% in 1L and 31% in 3L) and with at least 1 emergency room visit (27% in 1L and 41% in 3L). Mean total all-cause costs per-member per-month (PMPM) also increased with each subsequent LoT ($15,917 in 1L, $17,767 in 2L, and $18,266 in 3L). In addition, mean total all-cause costs of R/R FL patients defined as POD24 and double refractory were higher in 3L; $20,172 PMPM and $21,724 PMPM, respectively). Conclusions: In this elderly R/R FL population, similar therapies were used across multiple LoTs. With each subsequent LoT, treatment duration shortened and notably, costs and HCRU increased. Additionally, patients defined as POD24 and double-refractory experienced higher costs. This research highlights a substantial unmet need in R/R FL for novel therapeutic options that can provide better outcomes and reduce HCRU.

Validation of an Automated, Scalable Comprehensive Genomic Profiling Assay for Hematologic Malignancies

Introduction: The optimal diagnosis, prognosis, and treatment selection for hematologic disorders requires the assessment of somatic mutations across a subset of clinically relevant genes. Here we present the clinical validation results of a targeted next generation sequencing panel comprising all exons of 141 clinically relevant genes for myeloid and lymphoid malignancies. Methods: The panel was designed to include genes (n=141) based of clinical guidelines for myeloid diseases including acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), and myeloproliferative neoplasms (MPN), and lymphoid diseases including acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), and some lymphoma subtypes. The panel was validated to detect single nucleotide variants (SNVs), insertions/deletions (indels), FLT3 internal tandem duplications ( FLT3-ITDs), and sex. Gene level copy number alterations (CNAs) were validated for 11 genes and 5 sub-gene level CNAs in IKZF1, KMT2A, RUNX1, TET2, and TP53, and were selected based on clinical utility and frequency observed in a population of &amp;gt;10,000 microarray samples with confirmed myeloid or lymphoid indications. Custom hybridization probes (Twist Biosciences) were designed to interrogate all coding exons of the 141 genes using genomic libraries created from 250 ng of gDNA extracted from peripheral blood, bone marrow, or flow cytometry cell suspensions, followed by hybrid capture, and sequencing on Illumina DNA sequencers. Data analysis was performed using an in-house analysis pipeline to detect variants and perform rational filtering of technical artifacts. Validation studies were performed in a CAP/CLIA accredited clinical laboratory. Precision and concordance studies were performed using previously characterized clinical samples. SNVs, indels and FLT3-ITDs were orthogonally confirmed by an NGS-based laboratory developed test (ArcherDx) or by Sanger sequencing. CNAs were confirmed by digital multiplexed ligation-dependent probe amplification (dMLPA; MRC Holland). Results: Assay precision was determined using 15 previously characterized clinical samples with 36 SNVs with a variant allele frequency (VAF) range of 5.7-91.4%, 11 indels with a VAF range of 6.2-79.7%, one 24bp FLT3-ITD with a VAF of 30.2%, 17 deletions with a copy number (CN) range of 0.53-0.83, and 7 gains with CN range of 1.32-1.66. Three replicates of each sample were assessed for precision with 100% concordance observed for SNVs (36/36), indels (11/11), FLT3-ITD (1/1), CNAs (24/24), and gender (15/15). Inter-assay precision was assessed using multiple operators, instruments, reagent lots, and sample barcodes with the clinical samples. Interim analysis of concordance performed on 130 clinical samples demonstrated a positive percent agreement (PPA) of 99.7% for SNVs (334/335 in 114 clinical samples), 97.7% for indels (146/151 in 85 clinical samples), 98.7% for CNAs (30/30 in 17 clinical samples), and 96.7% for sex (127/130 in 130 clinical samples). Dilution series of both cell lines and clinical samples that will be used to determine analytical sensitivity are underway. Conclusions: This study highlights the clinical validation of a 141 gene NGS assay for the detection of clinically informative genomic alterations in hematologic malignancies. The detection of CNAs by NGS may be used to additionally confirm or monitor abnormalities previously detected by cytogenetics/microarray or inform the need for additional testing. Results from this validation study, when complete, will include at least 480 samples plus orthogonal testing. These data describe the performance of an assay to enable a comprehensive evaluation of genomic alterations using a single sample, further facilitating the use of broad NGS assays in hematologic malignancies.

Real-World Outcomes and Treatment Patterns in AL-Amyloidosis in Austria: An Analysis from the Austrian Interdisciplinary Amyloidosis Registry (AIDA)

Introduction This project aimed to provide an overview of the treatment landscape for AL - Amyloidosis (AL-A) in Austria as documented in AIDA, the Austrian Interdisciplinary Amyloidosis Registry. By examining the existing body of knowledge surrounding treatments and demographics, we seeked to shed light on the potential benefits and challenges for new treatment modalities in the context of AL-A. Methods We identified 210 AL- pts. from AIDA (cut off May 15, 2023). Statistics were descriptive. Kaplan Meier was chosen for survival plots and analyses were carried out with Excel and SPSS. Results We present data from 210 AL-A pts. recruited from AIDA. This constitutes 45.95% of all pts. in AIDA, with the major parts of the residual population made up by wt-ATTR amyloidosis pts. (47.48%), with some rare other amyloidosis subtypes accounting for the rest. There was a male pre-dominance in the cohort (62.38%) well in line with published data. Pts. with AL-A were found to be much younger (median age 66a vs. 78a) compared to pts. suffering from wt-ATTR. Diagnosis of AL-A was made between 1997 and 2023, with the median year of first diagnosis being 2016, resulting in median follow up in the cohort of ~ 7 years. The pattern of organ involvement by AL-A were quite typical with heart (78.6%), kidney (58.1%), GI-tract (23.3%) and PNS (21.4%) being the dominant manifestations. Advanced stages by either the classical Mayo staging, as well as the revised one were regrettably still by far more frequent at time of diagnosis as early disease stages (classical Mayo III in 65.8%, and revised MAYO III and IV in 27.8% and 42.3% of pts. respectively). This is mirrored in a high number of pts. with severe to moderate renal insufficiency (GFR &amp;lt; 30ml/min in 4.6% and between 30-60 ml/min in 42.2% of pts.) and/or moderate to severe cardiac involvement (NT-pro BNP &amp;gt; 5000 ng/l in 37.2%, and between 500 - 5000 ng/l in another 51.2% of pts.). Detailed data on comorbidities and toxicities will be added. Treatments Only 141/210 pts. ever received a documented 1 st line of treatment (LoT), of which only 35.5% received a 2 nd LoT, and even less a 3 rd LoT (20.6%), while more LoTs were applied only in a handful of pts. This horrendous attrition rate per LoT is far above, what has been described for Myeloma (20% / LoT). Time to next tretament was found to be median of 9.9 months. 54 pts. received a 1st LoT with the intent of pursuing an autologous stem cell transplantation (ASCT), but only 54% of these actually were transplanted. This compares inferior to Myeloma, although MM pts. are usually significantly older. The most frequently used therapeutic regimes were VCd-D (Andromeda) with or without Dara-maintenance. The widespread use of anti-CD38 based treatments in this sub-cohort is mirrored in a quite high rate of high-quality hematologic responses (CR+VGPR = 51.9%). 90 pts. received 1st LoTs not intending to lead to ASCT. Again, Daratumumab based regimes were the most used, but treatment outcomes were much inferior (hematologic CR+ VGPR = 18.9%) probably based on dosages, time on therapy, and not applied ASCT. Only 40 pts. received conventional 2nd LoTs usually classical Myeloma therapies, with modest treatment results. Even hematologic high-quality responses (CR+VGPR = 25%) were rare not to mention organ responses. Survival Median survival in the AIDA cohort was found to be 45 months. Broken down by Mayo stages, both classic and revised, was found to be quite unfavorable for advanced disease (14 months in revised Mayo stage IV or classical Mayo stage III). Conclusions Compared to other plasma cell dyscrasias overall outcomes in AL-A are still dismal, despite the introduction of recent effective anti-plasma cell therapeutics in the treatment algorithm. Disease modifying therapies are urgently needed and should be preferentially introduced in 1 st line, as only few pts. will receive 2 nd or later LoTs. Furthermore, efforts to improve awareness for this dreadful disease are urgently needed to enable more therapies to be applied in less advanced disease settings. Take home-messages: 1. Efforts needed to improve early diagnosis of AL-A when treatment options will not be hampered by progressive organ dysfunctions 2. Absolute necessity to explore disease modifying therapies going beyond stopping excess free light chain production 3. As only a limited number of pts. received multiple LoTs any new therapeutics should be integrated in effective 1 st LoTs preferentially.

648. Validation of a Synthetic Multiplexed External Control for Monitoring the Performance and Detection of Multiple Respiratory Viral and Bacterial Pathogens for a Point of Care Nucleic Acid Testing Panel

Abstract Background Advancements in nucleic acid testing has enabled point of care testing solutions to provide clinicians with greatly improved time to diagnosis and patient care. However, complex technologies and regulatory requirements demand close monitoring of assay performance to accurately identify shifts, trends, and random errors to ensure accurate patient reporting. To support these requirements, MMQCI has developed a synthetic, easy-to-use, external control that simultaneously monitors the accurate identification of 15 respiratory pathogens (11 viral, 4 bacterial targets) detected with the BIOFIRE® SPOTFIRE® Respiratory (R) Panel. Methods The multiplex control contains genome segments of all pathogens detected by the SPOTFIRE R Panel. Sequences were designed in silico, ligated into MMQCI vectors, and transformed to create stable frozen clone stocks. In vitro RNA transcripts were generated, quantified, and formulated in a proprietary matrix to stabilize and carry the RNA through the entire test process. Validation studies were conducted to demonstrate precision across multiple reagents, operators and sites. Failure-mode, shipping and stability studies were conducted to confirm robustness and reliability. Three unique lots of the SPOTFIRE RSP Positive and Negative controls were manufactured to include variability in key components and tested across 6 different sites, incorporating multiple reagent lots, operators and instruments. A total of 361 runs were performed across the positive and negative controls using the SPOTFIRE R Panel. Results Of the 361 total runs across the 6 sites, 351 were valid and used for analysis. A total of 171/172 positive controls resulted in correct calls (99.3%), and all 179 negative controls were called correctly (100%). A regression analysis was performed to confirm real-time stability for 12 months when stored at temperatures ranging from 2oC to 25oC.Table 1.Summary of Reproducibility Test Results: External and Internal Site Conclusion Validation studies demonstrated reproducibility, and overall precision and robustness of SPOTFIRE RSP Positive and Negative Controls. The SPOTFIRE RSP Positive and Negative Controls provide a stable, ease to use solution for training, verification and routine monitoring of the performance of the BIOFIRE® SPOTFIRE® Respiratory Solution. Disclosures Nithya Sivaji, MS, Maine Molecular Quality Controls Inc.: employer Emma Farrell, n/a, Maine Molecular Quality Controls Inc.: employer Tania R. Spenlinhauer, PhD, Maine Molecular Quality Controls Inc.: employer Joan Gordon, n/a, Maine Molecular Quality Controls, Inc: President|Maine Molecular Quality Controls, Inc: Ownership Interest Monica Cronin, n/a, bioMerieux: Stocks/Bonds Brad Graham, n/a, bioMerieiuux: Employee Jeffrey Bastar, n/a, bioMerieux: Employee|bioMerieux: Stocks/Bonds

Preclinical Efficacy of a Novel CD8-Targeted Fusosome for In Vivo CAR T Cell Therapy

SG299 is a CD8-targeted fusosome; a self-inactivating (SIN) lentiviral vector (LVV) pseudotyped with a CD8-targeted fusogen, which carries a transgene encoding a CD19-directed chimeric antigen receptor (CAR). We evaluated the preclinical efficacy of a representative CD8/CD19CAR fusosome comparable to SG299 for its ability to control tumor growth in vivo in an NSG mouse model engrafted with human CD19 positive tumor cells and human peripheral blood mononuclear cells (PBMC). We compared direct intravenous (IV) administration and ex vivo exposure of fusosome to resting T cells followed by IV injection of vector-cell mixtures, as has been proposed for extracorporeal delivery (ECD) methods. We have also evaluated the performance of multiple lots of fusosomes to assess consistency in manufacturing and have evaluated donor-to-donor variability in five PBMC donors in this model. To measure anti-tumor efficacy, immunodeficient NSG mice were challenged IV with NALM6 tumors expressing firefly luciferase (5E5 cells/mouse) on day 4. For direct IV, PBMC (1E7 cells/mouse) were injected IV on day -1 followed by CD8/CD19CAR fusosome (1E7 IU/mouse) on day 0. For ECD, PBMC were incubated with CD8/CD19CAR fusosome for 1 hr and the cell and fusosome mix were injected IV on day 1. Tumor growth was monitored by bioluminescence imaging over a 28-day period. In addition, we have investigated both pharmacokinetics of fusosome induced CAR T cells and NALM6 tumors by flow cytometry. CD8/CD19CAR fusosome demonstrated statistically significant control of NALM6 tumors by both direct IV and ECD routes of delivery. Kinetic analysis of peripheral blood demonstrated tumor control and in vivo CART cell generation were dose dependent. The peak CAR T cell response was at D14. CARs detected at all timepoints were CD8+ specific; no CD4 + CAR T cells were observed. In addition, all mice that received fusosome demonstrated protection from morbidity and mortality as measured by reduced weight loss and improved survival compared to control or tumor only mice. Lot-to-lot performance variability of fusosomes was minimal; three lots of fusosome that had similar physical and functional attributes showed efficient and equivalent tumor control with multiple PBMC donors. Finally, an assessment of donor-to-donor variability across five PBMC donors using a single lot of fusosome showed donor-specific variations in tumor control as measured by BLI but demonstrated a consistent protection from morbidity and mortality. Collectively, these studies demonstrate preclinical efficacy of our CD8/CD19CAR candidate, lot consistency and verify our ability to generate CD19-directed CAR T cells in vivo using a specifically targeted platform.

An All-Human Hepatic Culture System for Drug Development Applications.

Finding a long-term, human-relevant culture model for primary human hepatocytes (PHHs) for pharmacological and toxicological studies remains a challenge. Current in vitro model platforms are often inconvenient and complex, lack phenotypic stability over time, and do not support multiple PHH lots, lacking experimental reproducibility and flexibility. Here, we provide a detailed protocol for the thawing, plating, and maintenance of an all-human 2D+ hepatic system (TV2D+), which takes advantage of standard two-dimensional (2D) culture techniques and equipment while maintaining the longevity and phenotypic stability over time that typically accompany more complex three-dimensional (3D) systems. The results show attachment and percent plateability in TV2D+ as a function of PHH seeding density, as well as stable functionality for at least 2 weeks in culture. A range of PHH seeding densities are assessed to achieve a successful long-term culture. When established properly, the PHHs in TV2D+ organize into hepatocyte colonies, express a hepatic-specific marker, and maintain viability, architectural integrity, and physiologically relevant levels of albumin and urea. This unique combination of attributes makes the TV2D+ system a suitable hepatic model for a variety of pharmacological and toxicological applications.