Three-year efficacy, overall survival, and safety of ruxolitinib therapy
Transcription
Three-year efficacy, overall survival, and safety of ruxolitinib therapy
Published Ahead of Print on January 23, 2015, as doi:10.3324/haematol.2014.115840. Copyright 2015 Ferrata Storti Foundation. Three-year efficacy, overall survival, and safety of ruxolitinib therapy in patients with myelofibrosis from the COMFORT-I study by Srdan Verstovsek, Ruben A. Mesa, Jason Gotlib, Richard S. Levy, Vikas Gupta, John F. DiPersio, John V. Catalano, Michael W.N. Deininger, Carole B. Miller, Richard T. Silver, Moshe Talpaz, Elliott F. Winton, Jimmie H. Harvey, Murat O. Arcasoy, Elizabeth O. Hexner, Roger M. Lyons, Azra Raza, Kris Vaddi, William Sun, Wei Peng, Victor Sandor, and Hagop Kantarjian Haematologica 2015 [Epub ahead of print] Citation: Verstovsek S, Mesa RA, Gotlib J, Levy RS, Gupta V, DiPersio JF, Catalano JV, Deininger MW, Miller CB, Silver RT, Talpaz M, Winton EF, Harvey JH, Arcasoy MO, Hexner EO, Lyons RM, Raza A, Vaddi K, Sun W, Peng W, Sandor V, and Kantarjian H.Three-year efficacy, overall survival, and safety of ruxolitinib therapy in patients with myelofibrosis from the COMFORT-I study. Haematologica. 2015; 100:xxx doi:10.3324/haematol.2014.115840 Publisher's Disclaimer.0 E-publishing ahead of print is increasingly important for the rapid dissemination of science. Haematologica is, therefore, E-publishing PDF files of an early version of manuscripts that have completed a regular peer review and have been accepted for publication. E-publishing of this PDF file has been approved by the authors. After having E-published Ahead of Print, manuscripts will then undergo technical and English editing, typesetting, proof correction and be presented for the authors' final approval; the final version of the manuscript will then appear in print on a regular issue of the journal. All legal disclaimers that apply to the journal also pertain to this production process. Three-year efficacy, overall survival, and safety of ruxolitinib therapy in patients with myelofibrosis from the COMFORT-I study Srdan Verstovsek,1 Ruben A. Mesa,2 Jason Gotlib,3 Richard S. Levy,4 Vikas Gupta,5 John F. DiPersio,6 John V. Catalano,7 Michael W.N. Deininger,8* Carole B. Miller,9 Richard T. Silver,10 Moshe Talpaz,11 Elliott F. Winton,12 Jimmie H. Harvey,13 Murat O. Arcasoy,14 Elizabeth O. Hexner,15 Roger M. Lyons,16 Azra Raza,17 Kris Vaddi,4 William Sun,4 Wei Peng,4 Victor Sandor,4 and Hagop Kantarjian,1 for the COMFORT-I investigators 1 The University of Texas MD Anderson Cancer Center, Houston, TX, USA 2 Mayo Clinic, Scottsdale, AZ, USA 3 Stanford Cancer Institute, Stanford, CA, USA 4 Incyte Corporation, Wilmington, DE, USA 5 Princess Margaret Cancer Centre, University of Toronto, Toronto, ON, Canada 6 Washington University School of Medicine, St. Louis, MO, USA 7 Frankston Hospital and Department of Clinical Haematology, Monash University, Frankston, Australia 8* Oregon Health and Science University, Portland, OR, USA 9 Saint Agnes Cancer Institute, Baltimore, MD, USA 10 Weill Cornell Medical Center, New York, NY, USA 11 University of Michigan, Ann Arbor, MI, USA 12 Emory University School of Medicine, Atlanta, GA, USA 13 Birmingham Hematology and Oncology, Birmingham, AL, USA 14 Duke University Health System, Durham, NC, USA 15 Abramson Cancer Center at the University of Pennsylvania, Philadelphia, PA, USA 16 Cancer Care Centers of South Texas/US Oncology, San Antonio, TX, USA 17 Columbia Presbyterian Medical Center, New York, NY, USA *Currently at Division of Hematology and Hematologic Malignancies and Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA A complete list of the COMFORT-I investigators appears in the Appendix. Running head: COMFORT-I study: 3-yr update on efficacy and safety Correspondence: Srdan Verstovsek, MD, PhD; e-mail: [email protected] Current text word count: ~3300 Abstract word count: 200 Figure/table count: 6 figures/1 table; 1 supplemental figure/4 supplemental tables Supplemental files: 1 file Trial registration: clinicaltrials.gov identifier: NCT00952289 Funding COMFORT-I was supported by Incyte Corporation. Acknowledgments This research is supported in part by the MD Anderson Cancer Center Support Grant CA016672. The authors would like to thank the COMFORT-I investigators for their participation in this trial. The authors would also like to acknowledge the medical writing assistance of Alfredo Toschi, PhD, of Evidence Scientific Solutions, which was funded by Incyte Corporation. Key words: ruxolitinib, myelofibrosis, overall survival, splenomegaly ABSTRACT In the phase III COMFORT-I study, the Janus kinase 1 (JAK1)/JAK2 inhibitor ruxolitinib provided significant improvements in splenomegaly, key symptoms, and quality-of-life measures and was associated with an overall survival benefit relative to placebo in patients with intermediate-2 or high-risk myelofibrosis. This planned analysis assessed the long-term efficacy and safety of ruxolitinib at a median follow-up of 149 weeks. At data cutoff, approximately 50% of patients originally randomized to ruxolitinib remained on treatment whereas all patients originally assigned to placebo had discontinued or crossed over to ruxolitinib. At week 144, mean spleen volume reduction was 34% with ruxolitinib. Previously observed improvements in quality-of-life measures were sustained with longer-term ruxolitinib therapy. Overall survival continued to favor ruxolitinib despite the majority of placebo patients crossing over to ruxolitinib (hazard ratio 0.69 [95% confidence interval: 0.46-1.03]; P=0.067). Exploratory analyses suggest that crossover may have contributed to an underestimation of the true survival difference between the treatment groups. Ruxolitinib continued to be generally well tolerated; there was no pattern of worsening of grade ≥3 anemia or thrombocytopenia with longer-term ruxolitinib exposure. These longer-term data continue to support the efficacy and safety of ruxolitinib in patients with myelofibrosis. The study is registered at clinicaltrials.gov: NCT00952289. Introduction Myelofibrosis (MF) is a Philadelphia chromosome–negative myeloproliferative neoplasm (MPN) that may occur as primary myelofibrosis (PMF) or develop from progression of polycythemia vera or essential thrombocythemia (post-PV or post-ET MF, respectively).1 The clinical hallmarks of MF are splenomegaly, cytopenias, and debilitating symptoms associated with a hypercatabolic state and systemic inflammation.2,3 Allogeneic stem cell transplantation is the only potentially curative therapy for MF. However, this therapy is recommended only for a limited number of patients because of the high risk of treatment-related morbidity and mortality as well as the poor medical condition of patients.4 Dysregulation of the Janus kinase (JAK)–signal transducer and activator of transcription (STAT) pathway is central to the pathogenesis of MF.5-8 Approximately 65% of patients with PMF carry the JAK2V617F gain-of-function mutation9 and 5-10% carry mutations in the thrombopoietin receptor gene (MPL).10,11 Additional mutations leading to dysregulation of the JAK-STAT signaling pathway have been characterized in patients with PMF and other MPNs, suggesting a high degree of complexity and heterogeneity in disease pathogenesis.12,13 Recently, mutations in the CALR gene encoding calreticulin were detected in approximately 67%14 to 82%15 of patients with ET and in 80%15 to 88%14 of patients with PMF who did not have JAK2 or MPL mutations. The high frequency of CALR mutations in these patients, along with evidence linking aberrant calreticulin activity to JAK-STAT activation, supports a role for calreticulin in the pathogenesis of MPNs.14 Despite the range of mutations, the central role of the JAK-STAT pathway in MPNs has provided the rationale for the development of targeted therapies that inhibit JAK-STAT signaling.16,17 The oral JAK1 and JAK2 inhibitor ruxolitinib has been evaluated in two phase III clinical trials in patients with intermediate-2 or high-risk PMF (per International Prognostic Scoring System)18 or post-PV MF or post-ET MF (per 2008 World Health Organization criteria): the randomized, double-blind Controlled MyeloFibrosis Study with ORal JAK Inhibitor Treatment (COMFORT)-I19 study (www.clinicaltrials.gov: NCT00952289) and the randomized, open-label COMFORT-II20 study (www.clinicaltrials.gov: NCT00934544), which compared the effects of ruxolitinib with placebo or best available therapy (BAT), respectively. Both studies showed that ruxolitinib treatment significantly reduced splenomegaly and provided marked improvements in MF-related symptoms and quality-of-life (QOL) measures compared with controls, regardless of JAK2V617F mutational status.19,20 The clinical benefit and safety of ruxolitinib treatment in COMFORT-I and COMFORT-II have been maintained with subsequent longer-term follow-up.2123 As anticipated, the effect of JAK2 inhibition on hematopoiesis resulted in dose-dependent anemia and thrombocytopenia. The majority of these cytopenias occurred in the first 8-12 weeks of treatment, and they were generally manageable with dose reductions and/or red blood cell transfusions. Subsequently, mean platelet counts stabilized and mean hemoglobin levels gradually returned to a new steady state just below baseline levels.21-24 Additionally, longer-term follow-up of the COMFORT studies has shown that ruxolitinib treatment was associated with an overall survival advantage, despite the crossover design of these studies.19,21-23 The objective of the current analysis is to provide an update on the efficacy, overall survival, and safety of ruxolitinib in patients enrolled in COMFORT-I at a median follow-up of approximately 3 years (149 weeks). Methods Patients and study design Detailed inclusion and exclusion criteria have been previously described.19 Briefly, patients with intermediate-2 or high-risk PMF or post-PV MF or post-ET MF and splenomegaly were randomized 1:1 to receive ruxolitinib or placebo orally twice a day (BID). The starting dose of ruxolitinib was based on baseline platelet count: 15 mg BID or 20 mg BID for baseline platelet counts of 100-200 × 109/L or >200 × 109/L, respectively. Doses could be modified per protocol.19 Crossover from the placebo arm to ruxolitinib was allowed prior to the primary analysis based on defined criteria for worsening splenomegaly. Upon completion of the primary analysis, the study was unblinded and all remaining patients receiving placebo were allowed to crossover to ruxolitinib.19 Each participating site’s institutional review board approved the protocol. The study sponsor analyzed and interpreted the data in collaboration with the investigators. All authors had access to the aggregate data and any additional analyses upon request. The study was conducted in accordance with the International Conference on Harmonization guidelines for Good Clinical Practice. All patients provided written informed consent.19 Assessments Timing and methods of assessment of spleen volume, symptom burden, QOL measures, and adverse events, described previously,19 are detailed in the Supplemental Appendix. Statistical analysis This prospectively defined analysis was to occur when all patients either reached the 144-week assessment or discontinued from study. Changes from baseline in spleen volume and palpable spleen length were based on observed cases and summarized descriptively. Durability of spleen volume reduction was evaluated using the Kaplan-Meier method in patients who achieved a ≥35% reduction from baseline. Loss of a ≥35% spleen volume reduction was defined as the first <35% spleen volume reduction from baseline that was also a ≥25% increase from nadir. Overall survival was assessed using the Kaplan-Meier method for the intent-to-treat population with patients assessed per their original randomized treatment. Survival time was measured from study start to last known status of the patient and was not censored at time of discontinuation from randomized treatment. The Cox proportional hazards model and log-rank test were used to calculate the hazard ratio with 95% confidence interval (CI) and P-value, respectively. The incidence of new-onset or worsening grade ≥3 anemia and thrombocytopenia, and of new-onset or worsening all-grade and grade ≥3 nonhematologic adverse events, were calculated using the life table method. Additional details on safety assessments are discussed in the Supplemental Appendix. To better understand the effect of crossover to ruxolitinib on survival measurement, two exploratory analyses were performed. The first used the rank-preserving structural failure time (RPSFT) method, a statistical method used in oncology trials to adjust for possible crossover effect.25-27 The second analysis was a parametric statistical modeling of overall survival using the generalized Gamma distribution,28,29 which fitted a 3-parameter regression model to the observed survival data to calculate the corresponding hazard of death for patients originally randomized to ruxolitinib or placebo. Full details and description of the exploratory analyses are described in the Supplemental Appendix. Results Patient disposition At a median follow-up of 149 weeks (range 19-175 weeks), 77 of the 155 patients (49.7%) originally randomized to ruxolitinib were still receiving ruxolitinib therapy. A total of 111 of the 154 patients originally randomized to placebo crossed over to ruxolitinib therapy. Of these 111 patients, 57 (51.4%) patients were still receiving ruxolitinib therapy (Figure 1). In patients originally randomized to ruxolitinib, discontinuation rates estimated using the Kaplan-Meier method were 21% at year 1, 35% at year 2, and 51% at year 3. Reasons for discontinuation included disease progression (23.1%), adverse events (19.2%), death (19.2%), and withdrawal of consent (15.4%) (Figure 1). The median exposure to ruxolitinib was 145 weeks for patients originally randomized to ruxolitinib; for these patients, the mean dose of ruxolitinib remained stable after initial dose adjustments in the first 8-12 weeks of therapy (Figure 2). For patients originally randomized to placebo, the median exposure to placebo was 37 weeks. For patients who crossed over to ruxolitinib from placebo, the median time to crossover was 41 weeks. The median exposure to ruxolitinib for patients who crossed over was 105 weeks, which, at the time of this analysis, was nearly three times longer than their exposure to placebo. Efficacy Reductions in spleen size were durable with longer-term treatment with ruxolitinib. The mean percentage change from baseline in spleen volume was −31.6% at week 24 (median −33.0%) and −34.1% at week 144 (median −38.4%) (Figure 3A). The mean percentage change from baseline in palpable spleen length was −43.4% at week 24 (median −41.2%) and −49.4% at week 144 (median −50.0%) (Figure 3A). Assessment of palpable spleen response using the International Working Group for Myelofibrosis Research and Treatment (IWG-MRT) and European LeukemiaNet (ELN) consensus criteria,30 showed that a palpable spleen response was achieved in 31.6% of ruxolitinib-treated patients compared with 2.0% of patients in the placebo arm at week 24. At week 144, palpable spleen response was achieved in 24.5% of patients originally randomized to ruxolitinib. Fifty-nine percent of patients (91/155) originally randomized to ruxolitinib achieved a ≥35% reduction in spleen volume at any time during the study follow-up. The majority of patients achieved a ≥35% reduction from baseline in spleen volume by week 12, the time of the first spleen volume assessment; in these patients, the probability of maintaining a ≥35% spleen volume reduction for at least 132 weeks corresponded to 144 weeks on therapy. In this analysis, the probability of maintaining a ≥35% spleen volume reduction for at least 132 weeks was 0.53. Over the course of follow-up, over 80% of patients who achieved a ≥35% reduction in spleen volume maintained a reduction of at least 10% (Figure 3B), a reduction that has been shown to be associated with meaningful improvements in QOL and MF-related symptoms.21 Although the modified MF Symptom Assessment Form version 2.0 was only assessed through week 24, improvements in QOL measures as assessed by the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30 (EORTC QLQ-C30) were maintained with longer-term therapy, including improvements in global health status/QOL, fatigue, role functioning, and physical functioning scales (Figure 4). Overall survival At the time of this analysis, there were 42 deaths among patients randomized to ruxolitinib and 54 deaths among those randomized to placebo. A list of causes of death is provided in Supplemental Data Table 1. With median follow-ups of 149.1 and 149.3 weeks for the ruxolitinib and placebo arms, respectively, the hazard ratio for overall survival continued to favor patients originally randomized to ruxolitinib compared with those originally randomized to placebo (hazard ratio 0.69 [95% CI: 0.46-1.03]; P=0.067) (Figure 5A). Although the hazard ratio continued to favor ruxolitinib, the P-value no longer reached nominal significance at the P=0.05 level. Therefore, exploratory analyses were conducted to assess the potential impact of crossover and substantially longer exposure to ruxolitinib than placebo among patients originally randomized to placebo. The RPSFT method estimated a hazard ratio of 0.36 (95% CI: 0.2041.035) (Figure 5B). Exploratory modeling of survival using a generalized Gamma distribution showed that the hazard of death in patients originally randomized to placebo was initially higher than in those randomized to ruxolitinib. This hazard subsequently decreased over time, corresponding with an increased proportion of patients who crossed over to ruxolitinib (Supplemental Data Figure 1). Safety As expected given the role of JAK2 in erythropoietin and thrombopoietin signaling, anemia and thrombocytopenia were the most common adverse events observed with ruxolitinib therapy. The incidence of new-onset or worsening grade 3 or 4 anemia and thrombocytopenia were highest during the first 6 months of therapy and decreased with longer-term ruxolitinib treatment (Figure 6A). Consistent with this observation, mean hemoglobin levels and platelet counts in ruxolitinib-treated patients decreased in the first 8-12 weeks. Mean hemoglobin levels reached a nadir in this time frame, then subsequently increased to a new steady-state level by week 24 and remained stable through the course of longer-term follow-up. Mean platelet counts remained stable with longer-term follow-up after the initial decrease (Figure 6B). Since the last reported analysis at a median follow-up of 102 weeks,21 no additional patients have discontinued from the study for anemia or thrombocytopenia. The most common nonhematologic adverse events that occurred more frequently with ruxolitinib compared with placebo in the primary analysis were ecchymosis (18.7%), dizziness (14.8%), and headache (14.8%).19 When adjusted for exposure to ruxolitinib, the incidence of these adverse events as well as other nonhematologic events decreased with longer-term therapy (Table 1), as did rates of grade ≥3 adverse events (Supplemental Data Table 2). Urinary tract infections (UTIs) and herpes zoster were infections that occurred in patients receiving ruxolitinib during randomized treatment;31 however, no increase in incidence was noted with long-term ruxolitinib therapy. A comprehensive analysis of MedDRA preferred terms associated with these infections showed the incidence of UTI per the life table method was 10.5% (n=15) for 0-<12 months, 6.7% (n=7) for 12-<24 months, 7.7% (n=6) for 24-<36 months, and 6.0% (n=2) for ≥36 months in patients originally randomized to ruxolitinib. Two UTIs were grade ≥3, one occurring between months 12 and 24 and one occurring between months 24 and 36. The incidences of herpes zoster were 2.1% (n=3) for 0-<12 months, 3.5% (n=4) for 12-<24 months, 3.4% (n=3) for 24-<36 months, and 0% for ≥36 months in patients originally randomized to ruxolitinib; all herpes zoster infections were grade 1 or grade 2. No other opportunistic infections occurred with long-term ruxolitinib therapy. The overall pattern of adverse events observed after treatment interruption or discontinuation continued to support the absence of a specific withdrawal effect (Supplemental Data Tables 3-4). Four new cases of acute myeloid leukemia (AML) have been reported since the previous analysis (two in patients originally randomized to ruxolitinib; two in patients originally randomized to placebo who developed AML after crossover to ruxolitinib), for a total of eight cases since study initiation (four in patients originally randomized to ruxolitinib; four in patients originally randomized to placebo). The rate of leukemic transformation per person-year of ruxolitinib exposure was 0.0121/person-year and 0.0233/person-year in patients originally randomized to ruxolitinib or placebo, respectively. Discussion In this planned analysis of the COMFORT-I study with a median follow-up of 149 weeks, ruxolitinib treatment continued to be associated with durable reductions in spleen volume and improvements in QOL measures. Longer-term follow-up revealed a slight decline in QOL measures. However, this may be related to the well-described phenomenon of “response shift,” which reflects the changes in patients’ perspective on key QOL domains owing to repeated testing over the course of treatment.32 Despite this potential for response shift, the EORTC QLQ-C30 scales indicated that QOL was still improved relative to baseline with longer-term ruxolitinib treatment. The hazard ratio for overall survival continued to favor ruxolitinib compared with placebo despite the majority of patients having crossed over from placebo to ruxolitinib, although statistical significance was not maintained. Although the rate of discontinuation from randomized treatment was higher in the placebo group than the ruxolitinib group at the primary analysis,19 follow-up for overall survival was well balanced between the two treatment arms. The exposure to ruxolitinib in patients who crossed over from placebo was substantially longer than their exposure to placebo (105 weeks vs. 37 weeks for median exposure to ruxolitinib and placebo, respectively), thus confounding the comparison of overall survival between the two treatment groups in favor of the placebo arm. To understand the effect of crossover to active treatment in placebo-controlled studies, several statistical methods have been developed. The exploratory analysis of overall survival using the RPSFT showed that crossover from placebo may have led to an underestimation of overall survival difference. This is consistent with findings from other oncology trials using this method, where crossover to active treatment may also have led to an underestimation of the survival difference between placebo and active treatment.26,27 Consistent with the RPSFT analysis, the exploratory analysis using the generalized Gamma function showed that the probability of death in the placebo group was initially higher than in the original ruxolitinib-treated group, and that this probability decreased over time as patients originally assigned to placebo crossed over to receive ruxolitinib treatment. This finding is expected for a crossover trial in which the active treatment has a positive impact on survival.29 Although the specific mechanism underlying the prolonged survival observed in patients originally randomized to ruxolitinib in COMFORT-I is unknown, the reductions in spleen volume and improvements in functional status and QOL measures may have had a modulatory effect on the common causes of death not related to disease progression in patients with MF.18 Consistent with our findings, a separate report of the COMFORT-II study showed that long-term ruxolitinib therapy was associated with an overall survival advantage relative to BAT at 3 years of follow-up (hazard ratio 0.48 [95% CI: 0.28-0.85]; P=0.009).23 Similar to what was observed in COMFORT-I, this analysis is likely biased against ruxolitinib owing to patients crossing over from BAT. However, in COMFORT-II the confounding effect of crossover is less severe than in COMFORT-I because of the longer exposure to BAT prior to crossover to ruxolitinib (median time of follow-up at primary analysis: 52 weeks in COMFORT-II20 and 32 weeks in COMFORT-I19). Additionally, a prespecified analysis of overall survival from pooled data from COMFORT-I and COMFORT-II supports an overall survival benefit of ruxolitinib compared with controls (hazard ratio 0.65 [95% CI: 0.46-0.90]; P=0.01). Further exploratory RPSFT analysis of pooled survival data from the COMFORT studies suggests an underestimation of the survival difference between treatment groups because of the effect of crossover (RPSFT-corrected hazard ratio 0.29 [95% CI: 0.13-0.63]; P=0.01).33 In this 3-year update of COMFORT-I, ruxolitinib treatment demonstrated durable efficacy at doses that were stable over the course of long-term follow-up. Dose adjustments occurred primarily in the first 8-12 weeks of the study, particularly in patients with baseline platelet counts between 100 and 200 × 109/L who received a starting dose of 15 mg BID. By week 24, the median titrated dose was 10 mg BID for this subgroup of patients and 20 mg BID for those with baseline platelet count >200 × 109/L; doses stabilized with longer-term treatment.24 Overall, no unexpected safety or tolerability issues were reported with longer-term ruxolitinib treatment. As expected, anemia and thrombocytopenia mainly occurred early in the course of treatment, and there was no pattern of worsening of these events with longer-term exposure to ruxolitinib in patients who remained in the study. An additional analysis of the incidence of new-onset or worsening grade ≥3 anemia and thrombocytopenia that counted patients who experienced both grade 3 and 4 events in each grade yielded similar results.34 As previously noted, the initial increases in anemia and thrombocytopenia observed in the first 6 months of treatment and the subsequent decline in the incidence of these events was consistent with the timing of ruxolitinib dose adjustments. In ruxolitinib-treated patients, the rate of red blood cell transfusions increased in the first 8 weeks of treatment and later declined to levels similar to the placebo arm by week 36 and remained stable thereafter. This was consistent with the observed pattern of hemoglobin levels, which initially decreased and subsequently stabilized at a new steady state.24 Although cases of UTIs and herpes zoster infections were observed in patients randomized to ruxolitinib, the incidence of these infections did not increase with longer- term therapy. As previously described, systematic review of the pattern of adverse events observed after treatment interruption or discontinuation in this analysis fails to support a specific withdrawal syndrome other than return to baseline disease.19,21 Longer-term ruxolitinib treatment did not affect the risk of transformation to AML. The rates of leukemic transformation per person-year of ruxolitinib exposure in patients originally randomized to ruxolitinib (0.0121/person-year) and in those originally randomized to placebo after they crossed over to ruxolitinib (0.0233/person-year) showed no evidence of an increased risk of leukemic transformation when compared with the rate derived from a historical control population of 310 patients with MF (0.038/person-year).35 In summary, patients receiving ruxolitinib treatment for a median of 3 years in the COMFORT-I study maintained durable reductions in spleen volume and meaningful improvements in QOL measures. Overall survival continued to favor those patients originally randomized to ruxolitinib compared with those originally randomized to placebo despite the majority of those assigned to placebo crossing over to ruxolitinib treatment. This crossover may have contributed to an underestimation of the true survival difference between the two treatment arms. Ruxolitinib treatment continued to be generally well tolerated, and the incidence of newonset grade 3 or 4 anemia and thrombocytopenia decreased with longer-term therapy. Collectively, long-term analyses from COMFORT-I and COMFORT-II continue to support the sustained efficacy and safety of ruxolitinib and provide evidence to support a meaningful ability of ruxolitinib to improve overall survival in patients with MF, and possibly modify the course of the disease. Authorship and disclosures SV, RAM, and HK performed the research and contributed to concept design, data collection, and data interpretation. JG, VG, JFD, JVC, MWND, CBM, RTS, MT, EFW, JHH Jr, MOA, EOH, RML, and AR performed research and contributed to data collection and interpretation. RSL and VS contributed to research design and data interpretation. WS and WP performed statistical analyses. KV contributed to research design. All authors assisted with drafting the manuscript and/or critical revision of the content and approved the final manuscript submitted. SV has received research funding from Incyte Corporation, AstraZeneca, Lilly Oncology, Roche, Geron, NS Pharma, Bristol Myers Squibb, Novartis, Celgene, Infinity Pharmaceuticals, YM Biosciences, Gilead, Seattle Genetics, Promedior, and Cell Therapeutics Inc. RAM has received research funding from Genentech, Gilead, Incyte, Lilly, NS Pharma, Sanofi-Aventis, and YM Biosciences. JG has served on an advisory committee and has received research funding and travel reimbursement from Incyte Corporation. VG has served as a consultant/advisory board member for Incyte and Novartis, and has received research funding from Celgene, Incyte, and Novartis through his institution. MWND has served as a consultant/advisory board for Bristol Myers Squibb, Ariad, Incyte, and Novartis, and has received research funding from Celgene, Bristol Myers Squibb, Gilead, and Novartis through his institution. CBM has served as a consultant/advisory board for Incyte and Novartis, and has received research funding from Incyte and Novartis through her institution. RTS has served on an advisory committee for Incyte and Gilead Sciences and has received honoraria from both companies. He has received research funding from Incyte, NS Pharma, Lilly Oncology, Novartis, Promedior, and Inhibikase Corp through his institution. MT has served as a consultant/advisory board member for Bristol Myers Squibb, Ariad, Novartis, and Pfizer. EFW has served as a consultant/advisory board member for Incyte, and has received research funding from Incyte through his institution. MOA has received research funding from Incyte through his institution. RML has served as a consultant/advisory board for Gilead, and has received research funding from Incyte, Celgene, Amgen, Lilly, Seattle Genetics, Gilead, Ariad, GlaxoSmithKline, and Novartis through his institution. AR has served in a speakers bureau for Novartis and has received research funding from Oncova Inc. HK has received research funding from Ariad, Novartis, Bristol Myers Squibb, and Pfizer through his institution. RSL, KV, WS, WP, and VS are employees of Incyte Corporation and have equity ownership. JFD, JVC, JHH Jr, and EOH have no relevant financial relationship to disclose. Appendix COMFORT-I Investigators The following investigators contributed to the study (listed in alphabetical order by country): Australia: P Cannell, Royal Perth Hospital, Perth, WA; JV Catalano, Frankston Hospital and Department of Clinical Haematology, Monash University, Frankston, Victoria; BH Chong, St. George Hospital, Kogarah, NSW; P Coughlin, Monash University/Box Hill Hospital, Box Hill, Victoria; STS Durrant, Royal Brisbane and Women’s Hospital, Herston, Queensland; TE Gan, Monash Medical Centre, Clayton, Victoria; HC Lai, Townsville Hospital, Douglas, Queensland; MF Leahy, Fremantle Hospital and Health Service, Fremantle, WA; M Leyden, Maroondah Hospital, Ringwood East, Victoria; R Lindeman, Prince of Wales Hospital, Randwick, NSW; D Ma, St. Vincent’s Hospital, Darlinghurst, NSW; A Perkins, Haematology and Oncology Clinics of Australia, Milton, Queensland; AC Perkins, Princess Alexandra Hospital, Woolloongabba, Queensland; D Ross, Flinders Medical Centre, Bedford Park, SA; W Stevenson, Royal North Shore Hospital, St. Leonards, NSW. Canada: K Grewal, Eastern Health, St. John’s, NL; V Gupta, Princess Margaret Hospital, University of Toronto, Toronto, ON; K Howson-Jan, London Health Sciences Centre, London, ON; S Jackson, St. Paul’s Hospital, Vancouver, BC; C Shustik, Royal Victoria Hospital, Montreal, QC; R van der Jagt, Ottawa Hospital-General Campus, Ottawa, ON. United States: L Afrin, Hollings Cancer Center, Charleston, SC; LP Akard, Indiana Blood and Marrow Transplantation, LLC, Beech Grove, IN; MO Arcasoy, Duke University Medical Center, Durham, NC; E Atallah, Froedtert Hospital and Medical College of Wisconsin, Milwaukee, WI; J Altman, Northwestern Memorial Hospital, Chicago, IL; J Camoriano, Mayo Clinic Arizona, Scottsdale, AZ; TP Cescon, Berks Hematology Oncology Associates, West Reading, PA; CR Cogle, University of Florida, Gainesville, FL; R Collins, Jr, University of Texas Southwestern Medical Center, Dallas, TX; KH Dao, Oregon Health and Science University, Portland, OR; HJ Deeg, Fred Hutchinson Cancer Research Center, Seattle, WA; M Deininger, Oregon Health and Science University, Portland, OR; NJ DiBella, Rocky Mountain Cancer Centers, Aurora, CO; JF DiPersio, Washington University School of Medicine, St. Louis, MO; A Faitlowicz, University of California-Irvine Medical Center, Orange, CA; FA Fakih, Florida Pulmonary Research Institute, LLC, Winter Park, FL; R Frank, Norwalk Hospital, Norwalk, CT; NY Gabrail, Gabrail Cancer Center Research, Canton, OH; SL Goldberg, Hackensack University Medical Center, Hackensack, NJ; J Gotlib, Stanford Cancer Institute, Stanford, CA; HM Gross, Dayton Physicians, LLC, Dayton, OH; JH Harvey, Jr, Birmingham Hematology and Oncology Associates, LLC, Birmingham AL; RH Herzig, University of Louisville, Louisville, KY; E Hexner, Abramson Cancer Center at the University of Pennsylvania, Philadelphia, PA; CE Holmes, Vermont Cancer Center, Burlington, VT; E Ibrahim, Beaver Medical Group, Highland, CA; R Jacobson, Palm Beach Cancer Institute, West Palm Beach, FL; C Jamieson, Moores University of California-San Diego Cancer Center, La Jolla, CA; K Jamieson, University of Iowa Hospitals and Clinic, Iowa City, IA; CM Jones, Jones Clinic, PC, Germantown, TN; HM Kantarjian, University of Texas MD Anderson Cancer Center, Houston, TX; A Kassim, Vanderbilt Clinic, Nashville, TN; CM Kessler, Georgetown University Medical Center, Washington, DC; T Kindwall-Keller, University Hospitals Case Medical Center, Cleveland, OH; PPN Lee, Tower Cancer Research Foundation, Beverly Hills, CA; RM Lyons, Cancer Care Centers of South Texas/US Oncology, San Antonio, TX; R Marschke, Jr, Front Range Cancer Specialists, Fort Collins, CO; J Mascarenhas, Mount Sinai School of Medicine, New York, NY; E Meiri, Palm Beach Institute of Hematology and Oncology, Boynton Beach, FL; A Menter, Kaiser Permanente, Denver, CO; RA Mesa, Mayo Clinic-Arizona, Scottsdale, AZ; C Miller, St. Agnes HealthCare, Inc., Baltimore, MD; C O’Connell, University of Southern California, Los Angeles, CA; I Okazaki, Straub Clinic and Hospital, Honolulu, HI; R Orlowski, Carolina Oncology Specialists, PA, Hickory, NC; R Paquette, University of California-Los Angeles Medical Hematology and Oncology, Los Angeles, CA; VR Phooshkooru, Mid Dakota Clinic, PC, Bismarck, ND; B Powell, Wake Forest University Health Services, Winston-Salem, NC; JT Prchal, Huntsman Cancer Institute, Salt Lake City, UT; R Ramchandren, Karmanos Cancer Institute, Detroit, MI; F Rana, Shands Jacksonville Clinical Center, Jacksonville, FL; A Raza, Columbia University Medical Center, New York, NY; C Rivera, Mayo Clinic-Jacksonville, Jacksonville, FL; EA Sahovic, Western Pennsylvania Hospital, Pittsburgh, PA; M Scola, Carol G. Simon Cancer Center, Morristown, NJ; M Scouros, Houston Cancer Institute, PA, Houston, TX; M Sekeres, Cleveland Clinic, Cleveland, OH; J Shammo, Rush University Medical Center, Chicago, IL; RS Siegel, George Washington University, Washington, DC; RT Silver, Weill Cornell Medical Center, New York, NY; CP Spears, Sierra Hematology and Oncology, Sacramento, CA; M Talpaz, University of Michigan Medical Center, Ann Arbor, MI; M Tsai, Park Nicollet Institute, St. Louis Park, MN; S Verstovsek, University of Texas MD Anderson Cancer Center, Houston, TX; T Walters, Mountain States Tumor Institute, Boise, ID; RS Weiner, Arena Oncology Associates, PC, Lake Success, NY; EF Winton, Emory University Hospital, Atlanta, GA; SE Young, Somerset Hematology-Oncology Associates, Somerville, NJ; F Yunus, University of Tennessee Cancer Institute, Memphis, TN. References 1. Abdel-Wahab O, Levine RL. Primary myelofibrosis: update on definition, pathogenesis, and treatment. Annu Rev Med. 2009;60:233-245. 2. Mesa RA, Schwager S, Radia D, et al. The Myelofibrosis Symptom Assessment Form (MFSAF): an evidence-based brief inventory to measure quality of life and symptomatic response to treatment in myelofibrosis. Leuk Res. 2009;33(9):1199-1203. 3. Tefferi A. Myelofibrosis with myeloid metaplasia. N Engl J Med. 2000;342(17):12551265. 4. Gupta V, Hari P, Hoffman R. Allogeneic hematopoietic cell transplantation for myelofibrosis in the era of JAK inhibitors. Blood. 2012;120(7):1367-1379. 5. James C, Ugo V, Le Couedic JP, et al. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature. 2005;434(7037):1144-1148. 6. Kralovics R, Passamonti F, Buser AS, et al. A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med. 2005;352(17):1779-1790. 7. Baxter EJ, Scott LM, Campbell PJ, et al. Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. Lancet. 2005;365(9464):1054-1061. 8. Levine RL, Wadleigh M, Cools J, et al. Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. Cancer Cell. 2005;7(4):387-397. 9. Tefferi A, Vainchenker W. Myeloproliferative neoplasms: molecular pathophysiology, essential clinical understanding, and treatment strategies. J Clin Oncol. 2011;29(5):573582. 10. Pikman Y, Lee BH, Mercher T, et al. MPLW515L is a novel somatic activating mutation in myelofibrosis with myeloid metaplasia. PLoS Med. 2006;3(7):e270. 11. Rumi E, Pietra D, Guglielmelli P, et al. Acquired copy-neutral loss of heterozygosity of chromosome 1p as a molecular event associated with marrow fibrosis in MPL-mutated myeloproliferative neoplasms. Blood. 2013;121(21):4388-4395. 12. Rampal R, Al-Shahrour F, Abdel-Wahab O, et al. Integrated genomic analysis illustrates the central role of JAK-STAT pathway activation in myeloproliferative neoplasm pathogenesis. Blood. 2014;123(22):e123-133. 13. Vainchenker W, Delhommeau F, Constantinescu SN, Bernard OA. New mutations and pathogenesis of myeloproliferative neoplasms. Blood. 2011;118(7):1723-1735. 14. Klampfl T, Gisslinger H, Harutyunyan AS, et al. Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med. 2013;369(25):2379-2390. 15. Nangalia J, Massie CE, Baxter EJ, et al. Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2. N Engl J Med. 2013;369(25):2391-2405. 16. Passamonti F, Maffioli M, Caramazza D, Cazzola M. Myeloproliferative neoplasms: from JAK2 mutations discovery to JAK2 inhibitor therapies. Oncotarget. 2011;2(6):485-490. 17. Rampal R, Al-Shahrour F, Abdel-Wahab O, et al. Integrated genomic analysis illustrates the central role of JAK-STAT pathway activation in myeloproliferative neoplasm pathogenesis. Blood. 2014;123(22):e123-e133. 18. Cervantes F, Dupriez B, Pereira A, et al. New prognostic scoring system for primary myelofibrosis based on a study of the International Working Group for Myelofibrosis Research and Treatment. Blood. 2009;113(13):2895-2901. 19. Verstovsek S, Mesa RA, Gotlib J, et al. A double-blind, placebo-controlled trial of ruxolitinib for myelofibrosis. N Engl J Med. 2012;366(9):799-807. 20. Harrison C, Kiladjian JJ, Al-Ali HK, et al. JAK inhibition with ruxolitinib versus best available therapy for myelofibrosis. N Engl J Med. 2012;366(9):787-798. 21. Mesa RA, Gotlib J, Gupta V, et al. Effect of ruxolitinib therapy on myelofibrosis-related symptoms and other patient-reported outcomes in COMFORT-I: a randomized, doubleblind, placebo-controlled trial. J Clin Oncol. 2013;31(10):1285-1292. 22. Cervantes F, Kiladjian J-J, Niederwieser D, et al. Long-term efficacy, safety, and survival findings from COMFORT-II, a phase 3 study comparing ruxolitinib with best available therapy for the treatment of myelofibrosis. Blood. 2012;120(21):Abstract 801. 23. Cervantes F, Vannucchi AM, Kiladjian JJ, et al. Three-year efficacy, safety, and survival findings from COMFORT-II, a phase 3 study comparing ruxolitinib with best available therapy for myelofibrosis. Blood. 2013;122(25):4047-4053. 24. Verstovsek S, Gotlib J, Gupta V, et al. Management of cytopenias in patients with myelofibrosis treated with ruxolitinib and effect of dose modifications on efficacy outcomes. OncoTarget Ther. 2014;4(7):13-21. 25. Robins JM, Tsiatis A. Correcting for non-compliance in randomized trials using rank preserving structural failure time models. Commun Stat Theory Methods. 1991;20(8):2609-2631. 26. Demetri GD, Garrett CR, Schoffski P, et al. Complete longitudinal analyses of the randomized, placebo-controlled, phase III trial of sunitinib in patients with gastrointestinal stromal tumor following imatinib failure. Clin Cancer Res. 2012;18(11):3170-3179. 27. Sternberg CN, Hawkins RE, Wagstaff J, et al. A randomised, double-blind phase III study of pazopanib in patients with advanced and/or metastatic renal cell carcinoma: final overall survival results and safety update. Eur J Cancer. 2013;49(6):1287-1296. 28. Lawless JF. Statistical Models and Methods for Lifetime Data. Hoboken, NJ: WileyInterscience; 2002. 29. Yavari P, Abadi A, Amanpour F, Bajdik C. Applying conventional and saturated generalized gamma distributions in parametric survival analysis of breast cancer. Asian Pac J Cancer Prev. 2012;13(5):1829-1831. 30. Tefferi A, Cervantes F, Mesa R, et al. Revised response criteria for myelofibrosis: International Working Group-Myeloproliferative Neoplasms Research and Treatment (IWG-MRT) and European LeukemiaNet (ELN) consensus report. Blood. 2013;122(8):1395-1398. 31. Incyte Corporation. Jakafi (ruxolitinib) tablets [prescribing information]. Wilmington, DE: Incyte Corporation; 2014. 32. Hamidou Z, Dabakuyo TS, Bonnetain F. Impact of response shift on longitudinal qualityof-life assessment in cancer clinical trials. Expert Rev Pharmacoecon Outcomes Res. 2011;11(5):549-559. 33. Vannucchi AM, Kantarjian H, Kiladjian JJ, et al. A pooled overall survival analysis of the COMFORT studies: 2 randomized phase 3 trials of ruxolitinib for the treatment of myelofibrosis. Presented at: 55th ASH Annual Meeting and Exposition; December 8, 2013; New Orleans, LA; December 8, 2013. (Abstract 2820) 34. Verstovsek S, Mesa AR, Gotlib J, et al. Long-term outcomes of ruxolitinib therapy in patients with myelofibrosis: 3-year update from COMFORT-I. Blood. 2013;122(21):Abstract 396. 35. Verstovsek S, Kantarjian HM, Estrov Z, et al. Long-term outcomes of 107 patients with myelofibrosis receiving JAK1/JAK2 inhibitor ruxolitinib: survival advantage in comparison to matched historical controls. Blood. 2012;120(6):1202-1209. TABLES Table 1. Incidence of new-onset all-grade nonhematologic adverse events regardless of causality. Ruxolitinib 12-<24 24-<36 0-<12 months months months ≥36 months (n=155) (n=130) (n=103) (n=82) Fatigue 29.0 15.2 15.3 7.7 Diarrhea 27.8 6.7 10.8 3.9 Ecchymosis 21.2 10.4 5.7 0 Peripheral edema 21.3 8.4 12.6 0 Dyspnea 19.2 10.2 2.9 3.3 Dizziness 18.1 10.4 3.0 3.5 Pain in extremity 18.0 6.2 4.2 3.3 Headache 16.6 5.1 2.7 0 Nausea 16.6 6.8 5.1 5.9 Constipation 14.5 8.6 10.1 9.0 Abdominal pain 13.8 5.7 3.6 0 Insomnia 13.8 5.7 3.7 0 Vomiting 13.7 2.8 2.4 5.5 Pyrexia 13.5 7.3 8.5 2.9 Cough 13.1 13.3 4.0 6.0 Arthralgia 11.8 5.8 6.6 6.3 Upper respiratory tract 7.7 11.1 4.0 3.2 Incidence, % infection Percentage of patients for each event was based on the effective sample size of the time interval (number of patients at risk at the beginning of the interval minus half of the censored patients during the time interval). Adverse event is included if the incidence was >10% at any yearly interval. FIGURE LEGENDS Figure 1. Patient disposition. *For the placebo arm, there were three patients who were not evaluable for safety (N=151); these patients were excluded from the calculation of the percentage of patients who discontinued (40/151). †Patients in the placebo group could crossover to ruxolitinib prior to the primary analysis based on defined criteria for worsening splenomegaly. After the primary analysis was completed, the study was unblinded and all remaining patients receiving placebo were allowed to crossover to ruxolitinib. ‡The percentages of patients who discontinued for the reasons listed are based on the number of patients who discontinued within the treatment group and not on the total number of patients in the treatment group. BID: twice a day. Figure 2. Mean daily dose of ruxolitinib over time in patients originally randomized to ruxolitinib. BID: twice a day; SEM: standard error of the mean. Figure 3. (A) Percentage change in spleen size over time. Mean percentage change from baseline in spleen volume (left panel) and palpable spleen length (right panel). (B) Durability of spleen volume reduction in patients originally randomized to ruxolitinib. The probability of maintaining a spleen volume reduction in patients who achieved a ≥35% decrease in spleen volume over the course of the study is shown. Also shown is the probability of maintaining a ≥10% spleen volume reduction—a spleen volume reduction that has been shown to be associated with meaningful improvements in quality of life and myelofibrosis-related symptoms21—in patients who achieved a ≥35% decrease in spleen volume. Figure 4. Mean change in quality-of-life (QOL) measures assessed using the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30. Shown are global health status, two functional domains (role and physical functioning), and symptom scores for fatigue, a key symptom impacting QOL, in patients with myelofibrosis. Arrows indicate improvement. Figure 5. (A) Overall survival in the intent-to-treat population as assessed by the KaplanMeier method. (B) Overall survival in the intent-to-treat population as assessed by the rank-preserving structural fail time (RPSFT) method. CI: confidence interval: HR: hazard ratio. Figure 6. (A) Incidence of new-onset or worsening grade 3 or 4 anemia and thrombocytopenia over time. (B) Mean hemoglobin level and platelet count and hemoglobin level over time. *All patients receiving placebo at the primary analysis crossed over or discontinued within 3 months of the primary analysis; therefore, data for patients receiving placebo are shown for up to 6 months only. RUX: ruxolitinib; PBO: placebo. Randomized (N=309) Ruxolitinib 15 mg BID or 20 mg BID (n=155) Placebo BID* (n=154) Crossed over to ruxolitinib (n=111)† Discontinued‡ 78 (50.3%) Death Adverse event Consent withdrawn Disease progression Noncompliance (meds) Noncompliance (study) Other 15 (19.2%) 15 (19.2%) 12 (15.4%) 18 (23.1%) 1 (1.3%) 1 (1.3%) 16 (20.5%) 54 (48.6%) Death Adverse event Consent withdrawn Disease progression Noncompliance (meds) Other 11 (20.4%) 8 (14.8%) 11 (20.4%) 15 (27.8%) 2 (3.7%) 7 (13.0%) 40 (26.5%) Death Adverse event Consent withdrawn Disease progression Other Remain on treatment 77 (49.7%) 57 (51.4%) 0 7 (17.5%) 9 (22.5%) 7 (17.5%) 13 (32.5%) 4 (10.0%) 20 mg BID starting dose 15 mg BID starting dose Mean daily dose (mg, BID) ± SEM 25 20 15 10 5 0 0 No. of patients 20 mg BID 100 15 mg BID 55 8 16 24 98 49 32 40 48 93 35 56 64 72 80 Weeks 77 33 88 96 73 30 104 112 120 69 26 128 136 144 62 20 A Spleen volume Mean percentage change from baseline 0 n= 148 139 120 107 100 Placebo 84 73 132 107 35 –10 –20 –30 –40 –50 –60 B 12 24 48 72 96 120 144 Weeks Ruxolitinib 10 Mean percentage change from baseline Ruxolitinib 10 Palpable spleen length 0 n= 153 152 150 141 130 110 102 90 79 147 141 136 109 47 –10 –20 –30 –40 –50 –60 12 24 48 Placebo 4 8 12 24 48 72 96 120 144 Weeks 4 8 12 24 48 1.0 ≥10% reduction (n=91) Probability 0.8 ≥35% reduction (n=91) 0.6 0.4 0.2 0 0 8 No. of patients at risk ≥35% 91 86 reduction 16 24 32 40 48 56 64 72 80 88 96 104 112 120 128 136 144 27 25 23 1 1 Weeks from initial ≥35% spleen volume reduction 77 75 68 62 59 54 51 49 42 40 38 37 Mean change from baseline Global health status/QOL 20 15 10 5 0 –5 –10 –15 0 12 24 36 48 60 72 84 Weeks 96 108 120 132 144 Role functioning Mean change from baseline 25 20 15 10 5 0 –5 –10 –15 –20 –25 Mean change from baseline 0 12 24 36 48 60 72 84 Weeks 96 108 120 132 144 Fatigue 10 5 0 –5 –10 –15 –20 –25 –30 –35 0 12 24 36 60 72 Weeks 84 96 108 120 132 144 Physical functioning 15 Mean change from baseline 48 10 5 0 –5 –10 0 12 24 36 48 60 72 Weeks 84 96 108 120 132 144 Ruxolitinib Placebo A 1.0 Original ruxolitinib Probability 0.8 Original placebogruxolitinib 0.6 HR: 0.69 (95% CI: 0.46-1.03); P=0.067 0.4 No. of deaths: ruxolitinib = 42; placebo = 54 Median follow-up: 149 weeks 0.2 Percentage of at-risk placebo who crossed over or discontinued 0 0 4 13 22 35 54 73 88 97 99 100 100 100 100 100 100 100 100 100 100 100 100 100 8 16 24 32 40 48 56 64 72 80 88 96 104 112 120 128 136 144 152 160 168 176 Weeks No. of patients at risk Ruxolitinib 155 155 153 148 145 143 137 131 125 124 122 115 112 111 111 108 106 101 84 45 19 1 0 Placebo 68 38 28 8 0 B 154 153 149 144 134 129 119 114 107 105 100 100 95 92 88 85 82 79 1.0 Probability 0.8 Ruxolitinib 0.6 Placebog ruxolitinib 0.4 Placebo-RPSFT 0.2 HR: 0.36 (95% CI: 0.204-1.035) 0 0 8 16 24 32 40 48 56 64 72 80 88 96 Weeks 104 112 120 128 136 144 152 160 168 176 A Ruxolitinib grade 4 Placebo* grade 3 Placebo* grade 4 Anemia 50 45 45 40 40 35 28.4 30 25 20 15 11.5 10.7 10 5 3.1 4.0 3.4 4.8 0-<6 6-<12 4.2 2.9 12-<18 B 18-<24 Months 1.6 0 24-<30 0 2.1 30-<36 0 0 ≥36 15 8.7 105 100 95 90 24 36 48 60 72 84 Weeks 0.8 3.4 0 0-<6 1.6 1.6 0.9 0.9 1.1 1.0 6-<12 12-<18 18-<24 Months 96 108 120 132 144 No. of patients RUX 155 145 143 136 124 113 110 107 104 100 94 PBO 151 132 113 83 37 88 79 3.5 1.1 2.1 1.9 24-<30 30-<36 0 0 ≥36 Platelet count 370 Mean platelets, × 109/L Mean hemoglobin, g/L 20 Placebo 110 12 25 0 Hemoglobin 115 0 30 5 Ruxolitinib 85 35 10 0 0 Thrombocytopenia 50 Percentage of patients Percentage of patients Ruxolitinib grade 3 320 270 220 170 120 0 12 24 36 48 60 72 84 Weeks 96 108 120 132 144 No. of patients RUX 155 144 143 136 124 112 110 107 104 100 94 PBO 151 128 112 82 37 88 79 Verstovsek S, et al. Online Supplement Detailed methods Patients and study design Patients enrolled in COMFORT-I had intermediate-2 or high-risk primary myelofibrosis (MF) according to the International Prognostic Scoring System1 or post–polycythemia vera MF or post-essential thrombocythemia MF according to the 2008 World Health Organization criteria, had splenomegaly (palpable ≥5 cm below the left costal margin), and had a platelet count ≥100 × 109/L. Detailed inclusion and exclusion criteria have been previously described.2 Eligible patients were randomized 1:1 to ruxolitinib or placebo given orally twice a day (BID). The starting dose of ruxolitinib was based on baseline platelet count: 15 mg BID for a baseline platelet count between 100 and 200 × 109/L or 20 mg BID for patients with a baseline platelet count >200 × 109/L. Doses could be decreased for safety or increased to enhance efficacy, as specified by the study protocol.2 The primary analysis occurred when all patients had either completed the week 24 evaluation or discontinued from the study and half of those remaining in the study completed the week 36 visit. Patients in the placebo group could crossover to ruxolitinib prior to the primary analysis based on defined criteria for worsening splenomegaly. After the primary analysis was completed, the study was unblinded and all remaining patients receiving placebo were allowed to crossover to ruxolitinib.2 The protocol was designed by Incyte Corporation and approved by the institutional review board at each participating site. The study sponsor’s clinical and statistical teams analyzed and interpreted the data in collaboration with the investigators. All authors had access to the aggregate study data and any additional analyses upon request. The study was conducted in accordance with the International Conference on Harmonization guidelines for Good Clinical Practice. All patients provided written informed consent.2 1 Verstovsek S, et al. Assessments The primary endpoint was the proportion of patients achieving a ≥35% reduction from baseline in spleen volume at week 24, assessed by abdominal imaging (magnetic resonance imaging or computed tomography).2 Spleen volume was measured at baseline, weeks 12, 24, 36, 48, 60, 72, and every 24 weeks thereafter. Palpable spleen length was assessed at baseline and at each study visit. Symptom burden, assessed by the modified MF Symptom Assessment Form version 2.0 electronic diary,2,3 was measured up to week 24. Quality of life was evaluated using the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30 at baseline and each study visit. Adverse events were evaluated using the National Cancer Institute Common Terminology Criteria for Adverse Events version 3.0. Statistical analysis The current analysis is a prospectively defined analysis of efficacy and safety, the timing of which was prespecified to occur when all patients either reached the 144-week assessment or discontinued from the study. Changes from baseline in spleen volume and palpable spleen length were based on observed cases and summarized descriptively. Durability of spleen volume reduction was evaluated using the Kaplan-Meier method in patients who achieved a ≥35% reduction from baseline in spleen volume. Loss of a ≥35% spleen volume reduction was defined as the first <35% spleen volume reduction from baseline that was also a ≥25% increase from nadir. Overall survival, a prespecified secondary endpoint, was assessed using the Kaplan-Meier method for the intent-to-treat (ITT) population with patients assessed per their original randomized treatment regardless of subsequent crossover. Survival time was measured from study start to last known status of the patient and was not censored at time of discontinuation from randomized treatment. The Cox proportional hazards model and log-rank test were used to calculate the hazard ratio with 95% confidence interval and P-value, 2 Verstovsek S, et al. respectively. The Kaplan-Meier method was used to estimate discontinuation rates at years 1, 2, and 3 based on time to discontinuation in the ruxolitinib arm. The incidence (conditional probability of event) of new-onset or worsening grade ≥3 anemia and thrombocytopenia (based on laboratory data) and of new-onset or worsening all-grade and grade ≥3 nonhematologic adverse events were calculated using the life table method based on the time to first event censored at the date of last laboratory evaluation for anemia and thrombocytopenia and the earlier of discontinuation or date of data cutoff for nonhematologic adverse events. Because the majority of the anemia and thrombocytopenia events occurred early in the study, the incidence of new-onset or worsening grade 3 or 4 anemia or thrombocytopenia was assessed at 6-month intervals in patients originally randomized to ruxolitinib; the placebo group was included only in the first 6-month interval because all patients receiving placebo discontinued or crossed over to ruxolitinib within 3 months of the primary analysis. The incidence of nonhematologic events was assessed in yearly intervals for patients originally randomized to ruxolitinib. Per the life table method, the incidence of each adverse event was based on the effective sample size of the time interval, which was the number of patients at risk at the beginning of the interval minus half of the censored patients during the time interval. To better understand the effect of patients from the placebo arm crossing over to ruxolitinib treatment on survival measurement, two exploratory analyses were performed. The first exploratory analysis used the rank-preserving structural failure time (RPSFT) method, a statistical method used in oncology trials to adjust for possible crossover effect.4-6 This method adjusts for crossover in the placebo group by relating the portion of observed survival time after crossover to active treatment to a multiplicative coefficient that represents either the beneficial or the harmful effect of treatment on survival, and applies this coefficient to estimate survival times as if crossover in the placebo group had not occurred. The hazard ratio was estimated using Cox regression analysis of reconstructed survival times, and the 95% confidence interval was estimated using the bootstrap method. As the null hypothesis of the RPFST method was 3 Verstovsek S, et al. the original ITT analysis, the method does not alter the P-value from the original ITT analysis. Additional model description and implementation details, including re-censoring of the reconstructed survival time, are described by Robins and Tsiatis4 and Korhonen et al.7 In the second analysis, a parametric statistical modeling of overall survival using the generalized Gamma distribution was conducted;8,9 this involved fitting a three-parameter regression model to the observed survival data that resulted in a smooth curve representing an estimated survival function curve. The survival function curve was then visually compared with the Kaplan-Meier curve over the period used for the model to understand how well it reflected the actual data. The fitted model was subsequently used to calculate the corresponding hazard of death for patients originally randomized to ruxolitinib and those randomized to placebo. References 1. Cervantes F, Dupriez B, Pereira A, et al. New prognostic scoring system for primary myelofibrosis based on a study of the International Working Group for Myelofibrosis Research and Treatment. Blood. 2009;113(13):2895-2901. 2. Verstovsek S, Mesa RA, Gotlib J, et al. A double-blind, placebo-controlled trial of ruxolitinib for myelofibrosis. N Engl J Med. 2012;366(9):799-807. 3. Mesa RA, Gotlib J, Gupta V, et al. Effect of ruxolitinib therapy on myelofibrosis-related symptoms and other patient-reported outcomes in COMFORT-I: a randomized, doubleblind, placebo-controlled trial. J Clin Oncol. 2013;31(10):1285-1292. 4. Robins JM, Tsiatis A. Correcting for non-compliance in randomized trials using rank preserving structural failure time models. Commun Stat Theory Methods. 1991;20(8):2609-2631. 4 Verstovsek S, et al. 5. Demetri GD, Garrett CR, Schoffski P, et al. Complete longitudinal analyses of the randomized, placebo-controlled, phase III trial of sunitinib in patients with gastrointestinal stromal tumor following imatinib failure. Clin Cancer Res. 2012;18(11):3170-3179. 6. Sternberg CN, Hawkins RE, Wagstaff J, et al. A randomised, double-blind phase III study of pazopanib in patients with advanced and/or metastatic renal cell carcinoma: final overall survival results and safety update. Eur J Cancer. 2013;49(6):1287-1296. 7. Korhonen P, Zuber E, Branson M, et al. Correcting overall survival for the impact of crossover via a rank-preserving structural failure time (RPSFT) model in the RECORD-1 trial of everolimus in metastatic renal-cell carcinoma. J Biopharm Stat. 2012;22(6):12581271. 8. Lawless JF. Statistical Models and Methods for Lifetime Data. Hoboken, NJ: WileyInterscience; 2002. 9. Yavari P, Abadi A, Amanpour F, Bajdik C. Applying conventional and saturated generalized gamma distributions in parametric survival analysis of breast cancer. Asian Pac J Cancer Prev. 2012;13(5):1829-1831. 5 Verstovsek S, et al. Supplemental Data Table 1. Causes of death by randomized treatment allocation.* Cause of death, n Ruxolitinib (N=155) Acute leukemia Acute myeloid leukemia Placebo (N=154) 1 2 2 Acute myeloid leukemia progression 1 Anastomotic hemorrhage 1 Anemia, systemic 1 Cardiac arrest 1 Cardiac failure congestive 1 Cerebral hemorrhage 1 Completed suicide 1 Congestive heart failure resulting from pneumonia 1 Death 1 Disease progression 6 9 Disease progression and cardiac failure 1 Gastrointestinal hemorrhage 1 Graft versus host disease 1 Intestinal perforation 1 Intra-abdominal hemorrhage 1 Leukemia or underlying leukemia 1 Leukemic transformation 1 Muscular weakness 1 1 Metastatic colon cancer 1 Multi-organ failure 1 Myelodysplastic syndrome disease progression 1 Myelofibrosis 3 Myelofibrosis progression 1 3 6 Verstovsek S, et al. Myelofibrosis with possible transformation to acute myeloid leukemia and pneumonia 1 Myeloproliferative disease 1 Myocardial infarction 2 Non-small cell lung cancer metastatic 1 Pancreatic carcinoma 1 Pneumonia 1 Pneumonia, multi-organ failure 1 Pneumonia and septic shock 1 Renal failure 1 Respiratory failure 1 Road traffic accident Sepsis or septic shock 1 1 3 Shock hemorrhagic 3 1 Shock, respiratory and cardiac failure; hemorrhage following splenectomy 1 Splenic infarction 1 Splenic rupture 1 Staphylococcal infection 1 Subdural hematoma 1 Subdural hemorrhage 1 Surgical complications 1 1 Unknown 9 11 Total 42 54 *Causes of death were collected verbatim as reported during long-term follow-up, thus cause of death was not available for all patients. 7 Verstovsek S, et al. Supplemental Data Table 2. Incidence of new-onset grade 3 or 4 nonhematologic adverse events regardless of causality. Ruxolitinib 12-<24 24-<36 0-<12 months months months ≥36 months (n=155) (n=130) (n=103) (n=82) Fatigue 6.2 0.9 3.3 0 Pneumonia 5.6 3.6 3.5 0 Abdominal pain 4.2 0 3.2 0 Arthralgia 2.1 0 0 0 Diarrhea 2.1 0 0 0 Dyspnea 2.1 0.9 2.2 2.5 Pain in extremity 2.1 0 1.1 0 Hyperuricemia 1.4 0.9 0 2.5 Fall 1.4 0.9 0 0 GI hemorrhage 1.4 0.9 0 0 Septic shock 1.4 0 0 0 Muscular weakness 1.4 0 1.1 0 Hypoxia 1.4 0 2.2 0 Sepsis 0.7 1.7 2.2 0 Epistaxis 0.7 1.7 0 0 Renal failure acute 0.7 0.9 2.2 2.4 Abdominal pain upper 0.7 0 2.2 0 Myocardial infarction 0 0.9 0 4.8 Incidence, % Percentage of patients for each event was based on the effective sample size of the time interval (number of patients at risk at the beginning of the interval minus half of the censored patients during the time interval). Adverse event is included if the incidence was ≥2 patients at any yearly interval. GI: gastrointestinal. 8 Verstovsek S, et al. Supplemental Data Table 3. Summary of treatment-emergent (grade 3 or higher) and SAEs reported during study drug interruption. Ruxolitinib (N=59) Adverse event Grade ≥3 Any SAE Number (%) of patients with any adverse event 27 (45.8) 21 (35.6) Anemia 9 (15.3) 3 (5.1) Thrombocytopenia 5 (8.5) 0 Hemoglobin decreased 2 (3.4) 1 (1.7) Pneumonia 2 (3.4) 2 (3.4) Sepsis 2 (3.4) 1 (1.7) Disseminated intravascular coagulation 1 (1.7) 0 Neutropenia 1 (1.7) 0 Platelet count decreased 1 (1.7) 1 (1.7) Cardiac failure congestive 1 (1.7) 1 (1.7) Abdominal pain 1 (1.7) 1 (1.7) Gastrointestinal hemorrhage 1 (1.7) 1 (1.7) Nausea 1 (1.7) 0 Obturator hernia 1 (1.7) 1 (1.7) Esophageal varices hemorrhage 1 (1.7) 1 (1.7) Rectal hemorrhage 1 (1.7) 1 (1.7) Retroperitoneal hematoma 1 (1.7) 0 Vomiting 1 (1.7) 1 (1.7) Asthenia 1 (1.7) 0 Fatigue 1 (1.7) 0 Systemic inflammatory response syndrome 1 (1.7) 0 Diverticulitis 1 (1.7) 0 Lung infection 1 (1.7) 1 (1.7) Perirectal abscess 1 (1.7) 1 (1.7) Pseudomonas sepsis 1 (1.7) 1 (1.7) 9 Verstovsek S, et al. Urinary tract infection 1 (1.7) 1 (1.7) Fall 1 (1.7) 0 Post-procedural hemorrhage 1 (1.7) 1 (1.7) Tibia fracture 1 (1.7) 1 (1.7) Troponin increased 1 (1.7) 0 Bone pain 1 (1.7) 0 Acute myeloid leukemia 1 (1.7) 1 (1.7) Delirium 1 (1.7) 0 Renal failure acute 1 (1.7) 0 Dyspnea 1 (1.7) 1 (1.7) Pneumonia aspiration 1 (1.7) 0 Pneumonitis 1 (1.7) 1 (1.7) Pulmonary hypertension 1 (1.7) 0 Febrile neutropenia 0 1 (1.7) Diastolic dysfunction 0 1 (1.7) Pyrexia 0 1 (1.7) Urosepsis 0 1 (1.7) All patients receiving placebo at the primary analysis crossed over or discontinued within 3 months of the primary analysis; therefore, no additional data beyond what were previously reported (Verstovsek S, Mesa RA, Gotlib J, et al. Efficacy, safety and survival with ruxolitinib in patients with myelofibrosis: results of a median 2-year follow-up of COMFORT-I. Haematologica. 2013;98(12):1865-1871) are available for these patients. SAEs: serious adverse events. 10 Verstovsek S, et al. Supplemental Data Table 4. Summary of treatment-emergent (grade 3 or higher) and SAEs reported after study drug discontinuation. Ruxolitinib (N=78) Adverse event Grade ≥3 Any SAE Number (%) of patients with any adverse event 31 (39.7) 32 (41.0) Thrombocytopenia 6 (7.7) 2 (2.6) Pneumonia 3 (3.8) 4 (5.1) Renal failure acute 3 (3.8) 1 (1.3) Sepsis 3 (3.8) 3 (3.8) Abdominal pain 2 (2.6) 2 (2.6) Acute myeloid leukemia 2 (2.6) 3 (3.8) Disease progression 2 (2.6) 2 (2.6) Dyspnea 2 (2.6) 1 (1.3) Hypokalemia 2 (2.6) 0 Hypotension 2 (2.6) 1 (1.3) Myocardial infarction 2 (2.6) 2 (2.6) Splenic infarction 2 (2.6) 2 (2.6) Acute respiratory failure 1 (1.3) 1 (1.3) Anemia 1 (1.3) 1 (1.3) Anuria 1 (1.3) 0 Asthenia 1 (1.3) 1 (1.3) Atrial fibrillation 1 (1.3) 0 Cardiac arrest 1 (1.3) 0 Cardiac failure congestive 1 (1.3) 1 (1.3) Cholecystitis infective 1 (1.3) 1 (1.3) Clostridial infection 1 (1.3) 1 (1.3) Confusional state 1 (1.3) 0 Death 1 (1.3) 1 (1.3) Disseminated intravascular coagulation 1 (1.3) 0 11 Verstovsek S, et al. Edema 1 (1.3) 0 Epistaxis 1 (1.3) 0 Fatigue 1 (1.3) 1 (1.3) Febrile neutropenia 1 (1.3) 0 Hemoglobin decreased 1 (1.3) 0 Hepatosplenomegaly 1 (1.3) 1 (1.3) Hyperbilirubinemia 1 (1.3) 0 Hyperglycemia 1 (1.3) 0 Hypoxia 1 (1.3) 0 Lactic acidosis 1 (1.3) 0 Leukocytosis 1 (1.3) 0 Malnutrition 1 (1.3) 0 Muscular weakness 1 (1.3) 1 (1.3) Pancreatic carcinoma 1 (1.3) 1 (1.3) Platelet count increased 1 (1.3) 0 Portal vein thrombosis 1 (1.3) 0 Pulmonary edema 1 (1.3) 0 Pulmonary tuberculosis 1 (1.3) 1 (1.3) Pyrexia 1 (1.3) 3 (3.8) Renal failure 1 (1.3) 1 (1.3) Renal tubular necrosis 1 (1.3) 0 Respiratory failure 1 (1.3) 1 (1.3) Septic shock 1 (1.3) 1 (1.3) Splenic hemorrhage 1 (1.3) 1 (1.3) Subdural hematoma 1 (1.3) 1 (1.3) Subdural hemorrhage 1 (1.3) 1 (1.3) Supraventricular tachycardia 1 (1.3) 0 Transaminases increased 1 (1.3) 0 Transient ischemic attack 1 (1.3) 1 (1.3) 12 Verstovsek S, et al. Abdominal pain upper 0 1 (1.3) Cellulitis 0 1 (1.3) Dehydration 0 1 (1.3) Diarrhea 0 1 (1.3) Fall 0 1 (1.3) Pneumonia aspiration 0 1 (1.3) Postoperative wound infection 0 1 (1.3) All patients receiving placebo at the primary analysis crossed over or discontinued within 3 months of the primary analysis; therefore, no additional data beyond what were previously reported (Verstovsek S, Mesa RA, Gotlib J, et al. Efficacy, safety and survival with ruxolitinib in patients with myelofibrosis: results of a median 2-year follow-up of COMFORT-I. Haematologica. 2013;98(12):1865-1871) are available for these patients. SAEs: serious adverse events. 13 Verstovsek S, et al. Supplemental Data Figure 1. Generalized Gamma distribution–based model of overall survival and hazard of death. 14