1837 Serum Proteomic Analysis of Multiple Myeloma Subjects Treated With Daratumumab Monotherapy Tineke Casneuf, 1 Andrew Lysaght, 2 Clare LeFave, 3 Jaime Bald, 4 Brendan Weiss, 5 Niels W.C.J. van de Donk, 6 Henk M. Lokhorst, 6 Tahamtan Ahmadi, 4 A. Kate Sasser 4, * 1Janssen Research & Development, Beerse, Belgium; 2Immuneering Corporation, Cambridge, MA, USA; 3LabConnect, LLC, Seattle, WA, USA; 4Janssen Research & Development, LLC, Spring House, PA, USA; 5Division of Hematology/Oncology, Department of Medicine, Abramson Cancer Center and Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; 6Department of Hematology, VU University Medical Center, Amsterdam, The Netherlands. *Presenting author. – Site ID was determined to be significantly associated with ✦ Expression of interleukin (IL)-1ß was significantly lower in – PD-L1/CD274 is the ligand for the receptor PD-1; the ✦ Two proteins, VCAM1 and GSN, were identified by all TLR2 ICOS CD163 INTRODUCTION METHODS the variables of interest (eg, demographics, response responders compared to non-responders ( P = 0.046; –7 engagement of PD-1 on T cells leads to reduced T-cell 3 applied statistical tests (shown in red text in Table 1 ) P = 0.001 14 P = 0.007 P = 2.53!10 11.0 17 class, and sample time point; ≥7.37% variability explained; Figure 4B ) 13.0 function and increased regulatory T-cell development Daratumumab Study Designs Wald test P = 3.71×10 –9 ), and ComBat 7 was utilized to Table 1. Proteins Exhibiting Differential Expression – In the bone marrow microenvironment, high 13 12.5 correct for site ID effects to reduce the impact of site- n 10.5 PD-L1 Between Baseline Versus On-Treatment Samples by o i ✦ Daratumumab is a first-in-class, human anti-CD38 IgG1 ✦ GEN501 was a phase 1/2, open-label, multicenter study concentrations of IL-1ß elicit the release of IL-6, which s related effects within the data s 12.0 Statistical Test e 14 r 12 Responders Non-responders monoclonal antibody in clinical development across the conducted in 2 parts ( Figure 3 ) promotes myeloma cell survival and expansion p x multiple myeloma (MM) disease spectrum ✦ For GEN501 Part 2, paired baseline and on-study samples E 10.0 11.5 ● Wilcoxon ANOVA:Visit Friedman – Part 1 was a dose-escalation phase ✦ E-cadherin (CDH1) and P-cadherin (CDH3) were shown to 14 from 16 patients were run in 1 batch and the data were VCAM1 VCAM1 VCAM1 1,2 11 11.0 ✦ High levels of CD38 are expressed on myeloma cells be increased in responders ( P = 0.007 and 0.012, GSN GSN GSN – Part 2 was a dose-expansion phase pre-processed with the SomaLogic standardization 9.5 ● ● respectively; Figure 4B ) 10.5 ● CXCL12 THBS4 CXCL12 ✦ The antimyeloma activity of daratumumab is mediated workflow 13 ● ● IGHM MMP3 Baseline On BaselineOn Baseline On ● ● ● ✦ SIRIUS was a phase 2, open-label, international, multicenter ● treatment treatment treatment ● ● through a number of mechanisms of action, including – Soluble serum E-cadherin is an independent prognostic PRTN3 ● study of Simon 2-stage design ( Figure 3 ) ● ● ● n ● ● ● PLG Non-responder Responder Stable o complement-dependent cytotoxicity, antibody-dependent Statistical Analyses marker of poor survival for newly diagnosed patients i ● ● s ● s ● 12 ● THBS4 15 e ● cell-mediated toxicity, antibody-dependent cellular with MM r p ● ✦ Responders versus non-responders ● ● x MMP3 TLR2, toll-like receptor 2; ICOS, inducible T-cell co-stimulator; CD163, cluster of differentiation 163. ● 3,4 E ● ● ● phagocytosis, and apoptosis via cross-linking GEN501 SIRIUS ● ● 16 ● PIGR – The role of CDH3 is similar to that of CDH1 ● ● – Statistical comparison of protein concentration ● ● ● ● Boldface and underlined text indicates that the SOMAmer was also identified in the comparison of Figure 5. Immune-related proteins increased on ● ● ● 11 ● ● ✦ Two studies, GEN501 (ClinicalTrials.gov Identifier: Dose-escalation Randomization ● ● baseline versus on-treatment samples in SIRIUS. distributions in daratumumab responders (partial ● ● ● treatment in all response classes. ● VCAM1, vascular cell adhesion molecule 1; GSN, gelsolin; CXCL12, C-X-C motif chemokine 12; ● ● NCT00574288) and SIRIUS (NCT01985126), demonstrated A. ● IGHM, immunoglobulin heavy constant mu; PRTN3, proteinase 3; PLG, plasminogen; Color key ● ● response [PR], very good PR [VGPR], complete response ● ● THBS4, thrombospondin 4; MMP3, matrix metalloproteinase 3; PIGR, polymeric immunoglobulin and histogram ● that single-agent daratumumab had a tolerable safety ● receptor. Dose from 0.005-24 mg/kg 0 16 mg/kg 8 mg/kg [CR], and stringent CR [sCR]) versus non-responders 6 ● (n = 32) 10 0 (n = 16) (n = 18) t Differential Protein Changes in Responders Versus 4 profile and promising efficacy in patients with relapsed or n u ● (progressed disease [PD]) was performed at both baseline o 0 C refractory MM ( Figure 1 )5,6 2 Non-responders Due to Treatment and on treatment using 2 complementary methods: 0 Baseline On treatment Baseline On treatment Safety and response −4 −2 0 2 4 8 Row Z−Score ✦ 60 proteins were identified as having significantly evaluated Response evaluated (i) Wilcoxon rank-sum test on each individual SOMAmer Red lines indicate a decrease over time. Green lines indicate an increase over time. CONCLUSIONS 9 differential levels between responders and non-responders and (ii) Limma analysis on all SOMAmers simultaneously DYRK3 HMGB1 PD-L1, programmed cell death ligand 1. 40 PPIB An additional FLT3 ADAMTS13 upon daratumumab treatment CD163 35 TPM2 Dose-expansion 90 patients TNFSF8 – All P values were adjusted using the Benjamini-Hochberg CADM1 Figure 7. Changes in immune checkpoint marker PD-L1. ✦ Several proteins with differential expression CDC42BPB enrolled at DDR2 30 CAMK1 ENTPD5 ✦ Some of these proteins have known roles in myeloma DARA 16 mg/kg 10,11 MRC1 (BH) method for multiple hypothesis correction MST1 between responders and non-responders at baseline 25 BMP10 ADCYAP1 % TLR4.LY96 , biology (B-cell maturation antigen [BCMA], signaling IL1B 16 mg/kg 8 mg/kg 16 mg/kg CCL11 R were identified, many of which are associated with 20 KYNU GEN501 Part 2 CD84 R (n = 42) (n = 30) (n = 106) – Wilcoxon-adjusted P values are reported here CLEC7A HSD17B1 lymphocyte activation molecule [SLAM] family member 7 O 15 SIGLEC7 MM or CD38 HAPLN1 PSMA2 CSNK2A1 ✦ As in SIRIUS, treatment induced differential protein DNAJC19 [SLAMF7], transmembrane activator and CAML interactor 10 TNFRSF11A DARA, daratumumab. ✦ Baseline versus on treatment ING1 SHH CDC37 expression in GEN501 Part 2 ( Table 1 ) 5 IL11 [TACI], and beta-2-microglobulin [B2M]; Figure 6 ) ✦ 142 proteins were identified as differentially CNTF PES1 TNFSF9 0 CCL8 Figure 3. Study design for GEN501 and SIRIUS. – Baseline versus on treatment protein levels were TGM3 expressed between baseline and on treatment SOD2 CDH3 – Of the proteins identified, all but 3 were also identified in 8 mg/kg 16 mg/kg 8 mg/kg 16 mg/kg EGFR SERPINC1 MAP2K4 Immune and T-cell–related Proteins compared using 3 alternative statistical methods: (i) 2-way SERPINA4 GEN501 SIRIUS CDH1 NOTCH1 the comparison of baseline versus on-treatment samples – Two proteins involved in T-cell stimulation (TLR2 SEMA6A 12 SPARCL1 ✦ In both studies, patients were ≥18 years of age, had repeated-measures analysis of variance (ANOVA), (ii) the SET ✦ On treatment, expression of immune checkpoint marker HFE2 PDGFRB in SIRIUS PR VGPR CR sCR SLAMF7 and ICOS) increased in all patients 8 8 CFP R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R R documented myeloma requiring systemic therapy, and Wilcoxon signed-rank test, and (iii) the Friedman test R programmed cell death ligand 1 (PD-L1) decreased in N N N N N N N N N N N N N N N N N had an Eastern Cooperative Oncology Group performance responders and increased in non-responders compared to – These results further validate the observations in the ✦ 60 proteins were identified as differentially ORR, overall response rate; PR, partial response; VGPR, very good partial response; CR, complete – All P values were adjusted to control false discovery rate Centered and scaled protein concentrations are presented in the rows, and patients are presented response; sCR, stringent complete response. status of ≤2 in the columns. The horizontal color-bar located above the heat map indicates response class: baseline ( Figure 7 ) SIRIUS study expressed between daratumumab responders using the BH method for multiple hypothesis correction purple = non-responder, green = responder. Figure 1. Overall response rates with daratumumab versus non-responders over time; these proteins – In GEN501, patients had relapsed from or were refractory monotherapy in GEN501 and SIRIUS. – A 2-way repeated-measures ANOVA 12 was applied to B. include markers of tumor burden, which decrease in to ≥2 prior lines of therapy, including proteasome MST1 TNFSF8 TNFSF9 BCMA SLAMF7 determine if significant time-point:response-class 8.75 Responders Non-responders Responders Non-responders responders and increase in non-responders inhibitors (PIs), immunomodulatory drugs (IMiDs), P = 0.012 P = 0.003 P = 0.019 ● ● ● interaction occurred for each SOMAmer; a modified 13 12.0 ● ● SomaLogic Platform ● chemotherapy, and autologous stem cell transplantation ● ✦ On treatment, PD-L1 expression decreased in ● 18 ● 8.50 ● ● ● Wilcoxon rank-sum test was applied as a post hoc test to ● ● ● ● ● ● ● ● ● ● ● n ● responders and increased in non-responders ● ● ● ● ● ● ✦ The SOMAscan Assay (SomaLogic, Inc., Boulder,