Abstract: Idecabtagene vicleucel (ide-cel) was FDA-approved in March 2021 for the treatment of relapsed/refractory multiple myeloma after 4 lines of therapy. On the KarMMa trial, grade ≥ 3 cytopenias and infections were common. We sought to characterize cytopenias and infections within 100 days after ide-cel in the standard-of-care (SOC) setting. This multi-center retrospective study included 52 patients who received SOC ide-cel; 47 reached day-90 follow-up. Data were censored at day 100. Grade ≥ 3 cytopenia was present among 65% of patients at day 30 and 40% of patients at day 90. Granulocyte colony stimulating factor (G-CSF) was administered to 88%, packed red blood cell transfusions to 63%, platelet transfusions to 42%, thrombopoietin (TPO) agonists to 21%, intravenous immunoglobulin to 13%, and CD34+ stem cell boosts to 8%. At day 100, 19% and 13% of patients had ongoing use of TPO agonists and G-CSF, respectively. Infections occurred in 54% of patients and were grade ≥ 3 in 23%. Earlier infections in the first 30 days were typically bacterial (68%) and severe (50%). Later infections between days 31 and 100 were 50% bacterial and 42% viral; only 13% were grade ≥ 3. On univariate analysis, high pre-CAR-T marrow myeloma burden (≥ 50%), circulating plasma cells at pre-lymphodepletion (LD), and grade ≥ 3 anemia at pre-LD were associated with grade ≥ 3 cytopenia at both days 30 and 90. Longer time from last bridging treatment to LD was the only significant risk factor for infection.

Abstract: Multiple myeloma (MM) incidence, mortality, and survival vary by race and ethnicity, but the causes of differences remain unclear. We investigated demographic, clinical, and molecular features of diverse MM patients to elucidate mechanisms driving clinical disparities. This study included 495 MM patients (self-reported Hispanic, n = 45; non-Hispanic Black, n = 52; non-Hispanic White, n = 398). Hispanic and non-Hispanic Black individuals had an earlier age of onset than non-Hispanic White individuals (53 and 57 vs 63 years, respectively, P & lt; .001). There were no differences in treatment by race and ethnicity groups, but non-Hispanic Black patients had a longer time to hematopoietic cell transplant than non-Hispanic White patients (376 days vs 248 days; P = .01). Overall survival (OS) was improved for non-Hispanic Black compared with non-Hispanic White patients (HR, 0.50; 95% CI, 0.31-0.81; P = .005), although this association was attenuated after adjusting for clinical features (HR, 0.62; 95% CI, 0.37-1.03; P = .06). Tumor mutations in IRF4 were most common in Hispanic patients, and mutations in SP140, AUTS2, and SETD2 were most common in non-Hispanic Black patients. Differences in tumor expression of BCL7A, SPEF2, and ANKRD26 by race and ethnicity were observed. Clonal hematopoiesis was detected in 12% of patients and associated with inferior OS in non-Hispanic Black patients compared with patients without clonal hematopoiesis (HR, 4.36; 95% CI, 1.36-14.00). This study provides insight into differences in molecular features that may drive clinical disparities in MM patients receiving comparable treatment, with the novel inclusion of Hispanic individuals.

Abstract: Multiplex immunofluorescence (mIF) staining combined with quantitative digital image analysis is a novel and increasingly used technique that allows for the characterization of the tumor immune microenvironment (TIME). Generally, mIF data is used to examine the abundance of immune cells in the TIME; however, this does not capture spatial patterns of immune cells throughout the TIME, a metric increasingly recognized as important for prognosis. To address this gap, we developed an R package spatialTIME that enables spatial analysis of mIF data, as well as the iTIME web application that provides a robust but simplified user interface for describing both abundance and spatial architecture of the TIME. The spatialTIME package calculates univariate and bivariate spatial statistics (e.g. Ripley’s K, Besag’s L, Macron’s M and G or nearest neighbor distance) and creates publication quality plots for spatial organization of the cells in each tissue sample. The iTIME web application allows users to statistically compare the abundance measures with patient clinical features along with visualization of the TIME for one tissue sample at a time. Availability and implementation spatialTIME is implemented in R and can be downloaded from GitHub (https://github.com/FridleyLab/spatialTIME) or CRAN. An extensive vignette for using spatialTIME can also be found at https://cran.r-project.org/web/packages/spatialTIME/index.html. iTIME is implemented within a R Shiny application and can be accessed online (http://itime.moffitt.org/), with code available on GitHub (https://github.com/FridleyLab/iTIME). Supplementary information Supplementary data are available at Bioinformatics online.

Abstract: Myeloproliferative neoplasms (MPNs), including polycythemia vera, essential thrombocytosis, and primary myelofibrosis, are clonal hematopoietic neoplasms driven by mutationally activated signaling by the JAK2 tyrosine kinase. Although JAK2 inhibitors can improve MPN patients' quality of life, they do not induce complete remission as disease‐driving cells persistently survive therapy. ERK activation has been highlighted as contributing to JAK2 inhibitor persistent cell survival. As ERK is a component of signaling by activated RAS proteins and by JAK2 activation, we sought to inhibit RAS activation to enhance responses to JAK2 inhibition in preclinical MPN models. We found the SHP2 inhibitor RMC‐4550 significantly enhanced growth inhibition of MPN cell lines in combination with the JAK2 inhibitor ruxolitinib, effectively preventing ruxolitinib persistent growth, and the growth and viability of established ruxolitinib persistent cells remained sensitive to SHP2 inhibition. Both SHP2 and JAK2 inhibition diminished cellular RAS‐GTP levels, and their concomitant inhibition enhanced ERK inactivation and increased apoptosis. Inhibition of SHP2 inhibited the neoplastic growth of MPN patient hematopoietic progenitor cells and exhibited synergy with ruxolitinib. RMC‐4550 antagonized MPN phenotypes and increased survival of an MPN mouse model driven by MPL‐W515L. The combination of RMC‐4550 and ruxolitinib, which was safe and tolerated in healthy mice, further inhibited disease compared to ruxolitinib monotherapy, including extending survival. Given SHP2 inhibitors are undergoing clinical evaluation in patients with solid tumors, our preclinical findings suggest that SHP2 is a candidate therapeutic target with potential for rapid translation to clinical assessment to improve current targeted therapies for MPN patients.

Abstract: Background: Multiple myeloma (MM) is twice as common in Blacks compared to Non-Hispanic (NH) Whites and Hispanics. While treatment and mortality differences have been reported for Black patients with MM compared to NH White patients, there is limited data on Hispanic populations. Furthermore, the factors driving observed differences in MM presentation and treatment responses by race and ethnicity are largely unknown. We investigated demographic, clinical, and molecular features, including tumor mutations and clonal hematopoiesis (CH), in a diverse population of patients with MM to elucidate mechanisms driving clinical disparities. Methods: Patients diagnosed with MM who consented to our institutional biorepository protocol were eligible for inclusion. Demographic and clinical data were obtained from cancer registry and abstracted from electronic medical records. MM tumor cells were purified from bone marrow aspirates by CD138 affinity chromatography. DNA was isolated from tumor cells and whole blood for each patient, and whole exome sequencing (WES) data was generated. Tumor somatic mutations were characterized using paired tumor-normal (blood) WES. CH was classified based on blood-derived somatic mutations, using paired tumors and reference populations as germline comparators. Outcomes included overall survival (OS; date of diagnosis to death/last contact) and progression-free survival (PFS; 1 st-line treatment start to 1 st disease progression/death). Results: A diverse group of MM patients (n=496) were included: NH White (80%), NH Black (10%) and Hispanic (9%). NH Black and Hispanic MM patients had a younger median age at diagnosis (57 and 53 yrs, respectively) compared to NH Whites (63 yrs, p = 0.0001; Fig A). There was no statistical difference in treatment categories received by race/ethnicity. NH Black patients had a longer time to hematopoietic cell transplant (HCT; 376 days) than NH White or Hispanic patients (248 and 270 days, respectively, p = 0.011). There was an improvement in OS for NH Black (HR 0.49, 95% CI 0.30-0.81) and Hispanic (HR 0.66, 95% CI 0.37-1.18) patients compared to NH White patients, but the association was not statistically significant in Hispanics. In univariable analysis, OS was also associated with age at diagnosis, International Staging System (ISS), treatment with HCT, and treatment regimen category. In multivariable analysis, after adjusting for age, ISS, HCT, and treatment category there was no longer a statistically significant association between OS and race/ethnicity. Although a worse PFS was present among Hispanic patients (adjusted HR 1.45, 95% CI 0.99-2.13), there was no statistically significant difference in PFS by race/ethnicity. The most mutated genes in MM tumors were KRAS (24%), NRAS (17%), TP53 (11%), DIS3 (9%), and BRAF (9%) (Fig B). Genes with significantly higher tumor mutation rates in Black compared to NH White patients were SP140 (12% v 4%, p = 0.026), AUTS2 (8% v 2%, p = 0.04), and SETD2 (6% v 1%, p = 0.037). IRF4 was most commonly mutated in Hispanics (11% v 3% in NH White and 0% in Black, p = 0.019). We identified CH using WES in 60 (12%) patients. The most CH mutations were in ASXL1, DNMT3A, and TET2. There was no difference in the prevalence of CH by race/ethnicity (p=0.8). There was a statistically significant difference in OS by race/ethnicity and CH status (Fig C). For NH Black patients, CH (HR 4.36, 95% CI 1.36-14.0) and age at diagnosis (HR 1.08, 95% CI 1.03-1.14) were associated with inferior OS (Fig C). After adjusting for age in multivariable analysis, the positive association with CH status among Black patients was no longer statistically significant (HR 2.72, 95% CI 0.48-15.4). A positive, but not statistically significant, association for PFS in NH White patients with CH was also noted (adjusted HR 1.38, 95% CI 0.95-2.0). Conclusions: This is the first study to examine differences in tumor mutation profiles, CH, and treatment among different racial and ethnic groups of patients diagnosed with MM. Our data suggest that age at diagnosis, tumor mutations, and CH may all contribute to clinical disparities observed in patients with MM. Efforts to expand our cohort and incorporate additional molecular biomarkers, epidemiologic characteristics, and clinical parameters are ongoing with the ultimate goal of elucidating targetable biological mechanisms to personalize management and optimize outcomes for diverse patients diagnosed with MM. Figure 1 Figure 1. Disclosures Hampton: M2Gen: Current Employment. Blue: WebMD: Consultancy; Janssen Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees. Siqueira Silva: AbbVie Inc.: Research Funding; Karyopharm Therapeutics Inc.: Research Funding. Baz: GlaxoSmithKline: Consultancy, Honoraria; BMS, sanofi, Karyopharm, Janssen, AbbVie: Consultancy, Research Funding; Oncopeptides: Consultancy; Merck: Research Funding. Nishihori: Novartis: Research Funding; Karyopharm: Research Funding. Shain: Janssen oncology: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Karyopharm Therapeutics Inc.: Honoraria, Research Funding; Sanofi Genzyme: Consultancy, Speakers Bureau; Novartis Pharmaceuticals Corporation: Consultancy; GlaxoSmithLine, LLC: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; BMS: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Amgen Inc: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Adaptive Biotechnologies Corporation: Consultancy, Speakers Bureau; AbbVie: Membership on an entity's Board of Directors or advisory committees, Research Funding.

Abstract: Background: There are two first-line treatment recommendations for advanced ovarian cancer: i) upfront cytoreductive debulking followed by chemotherapy and ii) neoadjuvant chemotherapy prior to surgical debulking. The choice between these two treatment strategies is controversial as there are no standardized guidelines for clinical decision support. As such, there remains a critical unmet need to identity biomarkers to personalize the most effective treatment strategies. The primary objective of this study is to identify and validate radiomic biomarkers that predict treatment response among patients with high-grade serous ovarian cancer treated with upfront surgical debulking. Methods: Intratumoral radiomic features (n=308) were extracted from pre-treatment contrast-enhanced CT images; analyses were conducted to remove correlated, non-stable, and non-reproducible features. Patients treated with upfront surgery (N=182) were split into training (N=91) and test (N=91) cohorts and a cohort of patients treated with upfront neoadjuvant (N=116) was used to determine if the radiomic features were prognostic or predictive. Overall (OS) and progression-free survival (PFS) were the main endpoints. Classification and Regression Tree analysis was used to identity the most informative radiomic features in the training cohort, which were then analyzed in the test cohort and the upfront neoadjuvant cohort. Results: Decision tree analysis identified a volumetric feature, ROI volume center of mass (CoM) in X direction, as the most informative radiomic feature which stratified upfront surgery patients into high- and low-risk. In the training cohort, high-risk patients were associated with significantly worse OS versus low-risk patients (HR=2.01; 95% CI 1.07-3.77 vs. 1.00 and 5-year OS=39.9% vs. 56.1%, respectively; log-rank p-value=0.03). In the test cohort, the high- vs. low-risk patients were also associated with poor OS (HR=2.23; 95% CI 1.19-4.17 vs. 1.00 and 5-year OS=23.3% vs. 51.3%, respectively; log-rank p-value=0.01). This radiomic feature was not associated with OS among patients with upfront neoadjuvant (HR=1.23; 95% CI 0.73-2.07 vs. 1.00 and 5-year OS=23.9% vs. 36.6%, respectively; log-rank p-value=0.44). Similar findings were observed for PFS. Conclusion: Utilizing standard-of-care imaging, we identified and validated a predictive radiomic feature associated with outcomes among patients with high-grade serous ovarian cancer treated with upfront surgical debulking but not among patients treated with neoadjuvant chemotherapy. This radiomic biomarker, which describes the location of center of mass inside the tumor in pixels in the x-direction, could be potentially utilized as clinical decision support to guide first-line treatment options. This study was generously funded by a Miles for Moffitt pilot grant. Citation Format: Jaileene Perez-Morales, Christelle M. Colin Leitzinger, Sweta K. Sinha, Melissa J. McGettigan, Daniel K. Jeong, Olya Stringfield, Mahmoud Abdala, Natarajan Raghunand, Robert J. Gillies, Jing-Yi Chern, Lauren C. Peres, Matthew B. Schabath. Radiomic biomarkers to optimize treatment decision and predict patient outcomes in serous ovarian carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3218.

Abstract: A survival benefit has been consistently observed for tumor infiltrating lymphocytes (TILs) among ovarian cancer patients; however, prior studies consist of predominantly white women and little work has been conducted in racially diverse cohorts. Here, we investigate racial differences in the tumor immune landscape and survival among African-American (AA) and white women with high-grade serous ovarian carcinoma (HGSOC), the most common histotype of ovarian cancer. Leveraging two population-based case-control studies of ovarian cancer, the African-American Cancer Epidemiology Study and the North Carolina Ovarian Cancer Study, treatment-naïve AA women with HGSOC were matched to white women with HGSOC by stage and age. Multiplex immunofluorescence staining was performed on formalin-fixed paraffin-embedded whole tissue sections to measure TILs (CD3+) and T-cell subsets, cytotoxic (CD3+CD8+) and regulatory (CD3+FoxP3+) T-cells. Image analysis was completed on three regions of interest (ROI) selected from the intratumoral region. We categorized immune cell abundance within the tumor, stroma, and overall as & lt;1% and ≥1% positive cells. Multivariable Cox proportional hazard regression models were used to examine the association between immune cell abundance and survival overall and by race. Among 121 AA and 121 white women with HGSOC, more than half (56%) had a higher TIL infiltrate overall, while 33% and 11% had higher levels of cytotoxic and regulatory T-cells, respectively. No differences in immune cell abundance were observed by race. Mean follow-up time was 4.3 ± 5.2 years, and 72% of the women are deceased. Higher levels of TILs and cytotoxic T-cells were associated with better outcomes overall (hazard ratio [HR]=0.68, 95% confidence interval [CI] =0.53, 0.88 and HR=0.59, 95% CI=0.44, 0.80, respectively) and these associations were similar irrespective of tumor/stroma. No association with survival was observed for T-regulatory cells overall and in the tumor; however, improved survival was noted for higher levels of T-regulatory cells in the stroma (HR=0.69, 95% CI=0.49, 0.96). Associations with survival among white women were consistent with the overall findings, but among AA women, all associations were attenuated and not statistically significant. For example, white women with higher overall TILs had a 42% lower risk of all-cause mortality (HR=0.58, 95% CI=0.41, 0.82), whereas the association among AA women was weaker and not statistically significant (HR=0.79, 95% CI=0.54, 1.17). Adjusting for frontline treatment did not substantively impact these findings. Our results add to the existing evidence that a robust TIL infiltrate confers a survival advantage among women with HGSOC; however, AA women may not experience the same survival benefit as white women with HGSOC. An external replication in a larger cohort of ovarian cancer patients and additional investigation, particularly further characterization of the T-cells (e.g., exhaustion, activation) and their co-localization with other prognostically relevant immune cells, is warranted. Citation Format: Lauren C Peres, Christelle Colin-Leitzinger, Sweta Sinha, Jeffrey R. Marks, Jose R. Conejo-Garcia, Anthony J. Alberg, Elisa V. Bandera, Andrew Berchuck, Melissa L. Bondy, Brock C. Christensen, Michele L. Cote, Jennifer A. Doherty, Patricia G. Moorman, Carlos Moran Segura, Jonathan V. Nguyen, Edward S. Peters, Ann G. Schwartz, Paul D. Terry, Christopher M. Wilson, Brooke L. Fridley, Joellen M. Schildkraut. Racial differences in the tumor immune landscape and survival of high-grade serous ovarian carcinoma [abstract]. In: Proceedings of the AACR Virtual Conference: 14th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2021 Oct 6-8. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr PO-232.

Abstract: Tumor-infiltrating lymphocytes (TIL) confer a survival benefit among patients with ovarian cancer; however, little work has been conducted in racially diverse cohorts. Methods: The current study investigated racial differences in the tumor immune landscape and survival of age- and stage-matched non-Hispanic Black and non-Hispanic White women with high-grade serous ovarian carcinoma (HGSOC) enrolled in two population-based studies (n = 121 in each racial group). We measured TILs (CD3+), cytotoxic T cells (CD3+CD8+), regulatory T cells (CD3+FoxP3+), myeloid cells (CD11b+), and neutrophils (CD11b+CD15+) via multiplex immunofluorescence. Multivariable Cox proportional hazard regression was used to estimate the association between immune cell abundance and survival overall and by race. Results: Overall, higher levels of TILs, cytotoxic T cells, myeloid cells, and neutrophils were associated with better survival in the intratumoral and peritumoral region, irrespective of tissue compartment (tumor, stroma). Improved survival was noted for T-regulatory cells in the peritumoral region and in the stroma of the intratumoral region, but no association for intratumoral T-regulatory cells. Despite similar abundance of immune cells across racial groups, associations with survival among non-Hispanic White women were consistent with the overall findings, but among non-Hispanic Black women, most associations were attenuated and not statistically significant. Conclusions: Our results add to the existing evidence that a robust immune infiltrate confers a survival advantage among women with HGSOC; however, non-Hispanic Black women may not experience the same survival benefit as non-Hispanic White women with HGSOC. Impact: This study contributes to our understanding of the immunoepidemiology of HGSOC in diverse populations.