Citations

Abstract Expression of DLL3 was examined in additional tumor types, as it was found to be highly expressed in tumor-initiating cells (TIC) in small cell lung cancer (SCLC), where a DLL3-targeted antibody drug conjugate (ADC), rovalpituzumab tesirine (Rova-T; SC16LD6.5; Saunders et al. 2015 Sci Transl Med 7:302ra136)) exerted clinically meaningful anti-tumor effects in a phase I trial (Spigel et al. 2016 Lancet Oncology; In Press). DLL3 expression was profiled by qRT-PCR, ELISA and immunohistochemistry in multiple tumor types. Patient-derived xenografts (PDX) from melanoma and ovarian small cell carcinoma were established and used for efficacy studies to determine the ability of Rova-T to impact tumor growth and TIC frequency. DLL3 expression was seen in metastatic melanoma (55%), low grade gliomas (90%), glioblastoma (70%), medullary thyroid cancer (65%), carcinoids (33%), dispersed neuroendocrine tumors in the pancreas (9%), bladder (57%) and prostate (24%), testicular cancer (90%), and lung adenocarcinomas with neuroendocrine features (80%). Unlike SCLC, where DLL3 does not predict clinical outcome on standard therapies, DLL3 expression negatively correlates with overall survival in melanoma and small cell bladder cancer. In mice bearing DLL3 positive melanoma PDX, treatment with a single dose of Rova-T resulted in effective and durable responses (>100 days), which correlated with a significant impact on TIC frequency. Similarly, in mice bearing DLL3-positive ovarian small cell PDX, a single dose of Rova-T resulted in effective and durable responses (>100 days). Our results show that DLL3 is expressed in many neuroendocrine tumors (lung, ovarian, prostate, bladder, etc), metastatic melanoma, medullary thyroid cancer, low-grade gliomas and glioblastoma. Given pre-clinical results showing efficacy of Rova-T in melanoma and ovarian small cell carcinoma, as well as encouraging clinical data with Rova-T in patients with recurrent/refractory SCLC, clinical evaluation of Rova-T in DLL3-positive melanoma, glioblastoma, medullary thyroid cancer and other high-grade neuroendocrine carcinomas is warranted. A “basket” trial enrolling patients with DLL3-positive solid tumors is now recruiting patients (NCT02709889). Citation Format: Laura R. Saunders, Samuel A. Williams, Sheila Bheddah, Kumiko Isse, Sarah Fong, Marybeth A. Pysz, Himisha Beltran, Loredana Puca, Verena Sailer, Juan M. Mosquera, Yu Yin, Jiaoti Huang, Andrew J. Armstrong, Jorge Garcia, Cristina Magi-Galluzzi, Vadim Koshkin, Petros Grivas, Farhad Kosari, John Cheville, Justin C. Moser, Thomas J. Flotte, Thorvardur Halfdanarson, Aaron Mansfield, Konstantinos N. Leventakos, Julian R. Molina, Douglas W. Ball, Barry D. Nelkin, Jill E. Shea, Courtney L. Scaife, Scott J. Dylla. Expression of DLL3 in metastatic melanoma, glioblastoma and high-grade extrapulmonary neuroendocrine carcinomas as potential indications for rovalpituzumab tesirine (Rova-T; SC16LD6.5), a delta-like protein 3 (DLL3)-targeted antibody drug conjugate (ADC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3093. doi:10.1158/1538-7445.AM2017-3093

Abstract Background: The paucity of effective therapeutic agents for hepatocellular cancer (HCC) underscores the critical need for more effective therapeutic strategies. Recent studies indicate lipid biosynthesis and desaturation is required for HCC survival. Targeting these may prove beneficial because such changes contribute to therapeutic resistance. Stearoyl CoA desaturase (SCD1), a key mediator of fatty acid (FA) biosynthesis and rate-limiting in conversion of saturated fatty acids (SFAs) to mono-unsaturated fatty acids (MUFAs), is upregulated in HCC and many other cancers. As such, we therapeutically targeted a novel lipogenic tumor survival mechanism mediated by SCD1 as a means to combat the chemoresistance associated with HCC. In so doing, we evaluated a novel lead SCD1 inhibitor in HCC. Methods: Paraffin embedded patient HCC tissues were examined for SCD1 expression. Using combined computational and synthetic chemistry approaches, we synthesized four novel specific SCD1 inhibitors with SSI-4 being the lead SCD1 inhibitor. HCC cell lines were examined using proliferation assays for response to SSI-4. IC50 concentrations for blocking SCD1 enzyme activity was determined. Blood half-life and bioavailability of single dose SSI-4 was determined. Mechanisms of action of SCD1 were examined that included Endoplasmic reticulum (ER) stress. In vivo, antitumor activity was determined using HCC patient derived xenograft (PDX) mouse models. Results: We identified elevated SCD1 mRNA and protein in HCCs tissues. SSI-4 dose-dependently inhibits cell proliferation in HCC cell lines with specificity demonstrated by oleic acid (MUFA) co-culture. Single dose oral gavage SSI-4 demonstrated a half-life of ~4 hours and excellent oral bioavailability. SSI-4 was well tolerated with long-term daily dosing. SSI-4 treatment of HCC cells and tumors led to endoplasmic reticulum (ER) stress followed by apoptotic cell death. Single agent SSI-4 demonstrated antitumor activity in HCC PDX mouse models with suppression of ER stress regulated proteins. Conclusions: Targeting a novel lipid metabolic pathway in HCC may provide effective therapy for aggressive HCC. Citation Format: John Alton Copland, Laura A. Marlow, Ilah Bok, James L. Miller, Matsuda Akiko, Yan W. Asmann, Vivekananda Sarangi, Steven R. Alberts, Kabir Mody, Lewis R. Roberts, Mark J. Truty, Tushar C. Patel. Targeting stearoyl CoA desaturase 1 (SCD1) in hepatobilliary carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 192. doi:10.1158/1538-7445.AM2017-192

Objectives: The incidence of second anterior cruciate ligament (ACL) injury after ACL reconstruction (ACLR) and return to sport (RTS) in a young, active population is likely between 25-33% with the greatest risk in the first 12 months after RTS. Although the use of allograft tissue in young athletes has been reported to result in increased risk of graft failure, differences in graft and contralateral ACL injury risk between patients who receive a hamstrings (HS) autograft and bone-patellar tendon- bone (BTB) autograft has yet to be reported in a young, athletic population. The tested hypothesis was that the relative risk (RR) of ipsilateral graft failure would be higher in young, active patients who receive an ACLR with a hamstrings graft, while the RR of contralateral ACL injury would be higher in patients who receive and ACLR with a BTB autograft. Methods: One hundred thirty-nine subjects (99 female/40 male) with a mean age of 16.9±2.0 years old (range: 13-25 y/o) underwent ACLR with either a HS autograft (n=79) or a BTB autograft (n=60) and were released to return to pivoting/cutting sport. These patients were enrolled in a prospective, observational cohort study and were tracked for incidence of 2 nd ACL after ACLR for a median of 73 months. Thirty-four (24.5%) suffered a 2 nd ACL injury. Fisher’s exact tests were used to determine whether graft choice (HS vs. BTB) was associated with risk of either ipsilateral graft failure or contralateral ACL injury. Sub-group analyses by sex were also conducted. Results: Patients who underwent ACLR with HS graft (n=79) sustained 14 ipsilateral tears and 6 contralateral ACL injuries. Patients who received an ACLR with BTB (n=62) sustained 2 ipsilateral tears and 12 contralateral ACL injuries (Table 1). Patients who received an ACLR with HS graft were 6 times more likely (RR=6.2; 95% CI: 1.4-28.7) to suffer a graft failure after RTS than the BTB group. In the first 12 months after RTS, the HS graft patients were nearly 10 times (RR=9.5; 95% CI: 1.2-76.1) more likely to suffer an ipsilateral graft failure. Patients with a HS graft were 3 times less likely (RR=0.33; 95%CI: 0.12-0.935) to suffer a contralateral ACL injury and nearly 7 times less likely (RR=0.15, 95%CI: 0.031-0.709) to suffer a contralateral ACL injury in the first 12 months after RTS compared to the BTB group. When the cohort was divided by sex, females with HS grafts were significantly more likely to sustain a graft failure in the first 12 months after RTS (p=0.008) and by final follow-up (p=0.002) compared to females with BTB graft. Females with HS grafts were 3 times less likely (RR=0.30; 95%CI: 0.095-0.943) to suffer a contralateral ACL injury than females with BTB graft and 6 times less likely (RR=0.17; 95%CI: 0.034-0.846) to suffer contralateral ACL injury in the first year after RTS. Males demonstrated no significant difference in ipsilateral or contralateral injury risk based on graft type. Conclusion: Young, active females who return to pivoting and cutting sports after ACLR with HS autograft are at greater risk to suffer a graft failure and reduced risk to suffer a contralateral ACL injury compared to females who undergo ACLR with BTB graft tissue. No significant differences in 2 nd ACL injury risk based on graft type were observed in the male patients. [Table: see text]

Background: Previous work shows that young athletes after anterior cruciate ligament reconstruction (ACLR) demonstrate single-leg (SL) landing movement asymmetries at the time of return to sport (RTS); however, the effect of movement asymmetries on longitudinal knee-related function after ACLR has not been examined. Hypothesis/Purpose: The purpose of this study was to examine the effect of SL drop-landing movement symmetry at the time of RTS on knee-related function 2 years later in young athletes after ACLR. The first hypothesis was that young athletes who demonstrated SL drop-landing asymmetries at RTS would demonstrate decreased knee function 2 years later compared with those who demonstrated symmetric SL drop-landing mechanics. The second hypothesis was that SL drop-landing movement symmetry at RTS would be associated with knee functional recovery 2 years later. Study Design: Cohort study; Level of evidence, 2. Methods: This study included 48 young athletes who had undergone ACLR and were assessed at the time of RTS (77% female; mean [±SD] age at RTS, 17.6 ± 2.6 years) and followed for 2 years after RTS. Three sagittal-plane landing variables of interest were calculated using 3-dimensional motion analysis during an SL drop-landing task at the time of RTS: knee flexion excursion, peak internal knee extension moment, and peak trunk flexion. The limb symmetry index (LSI) was calculated for each landing variable using the following: LSI = (involved/uninvolved) × 100%. The LSI was used to divide the cohort into symmetric (SYM) and asymmetric (ASYM) groups for each landing variable: knee flexion excursion (SYM: LSI ≥ 90% [n = 23]; ASYM: LSI < 85% [n = 18]), peak internal knee extension moment (SYM: LSI ≥ 90% [n = 19]; ASYM: LSI < 85% [n = 22]), and peak trunk flexion (SYM: LSI ≤ 105% [n = 25]; ASYM: LSI > 115% [n = 19]). At 2 years after RTS, knee-related function was evaluated using the Knee Injury and Osteoarthritis Outcome Score (KOOS), International Knee Documentation Committee (IKDC) subjective knee form, and performance on SL hop tests. Functional recovery was defined based on literature cutoffs for knee-related functional measures. Differences in 2-year function were compared between the symmetry groups using Mann-Whitney U tests because of nonnormality. Logistic regression was used to determine if landing symmetry at the time of RTS would be associated with 2-year knee functional recovery after RTS. Results: The ASYM knee flexion excursion group demonstrated decreased function at 2 years after RTS compared with the SYM group on the KOOS-Pain (ASYM: 93.0 ± 8.2; SYM: 98.4 ± 3.0; P = .008) and the KOOS–Quality of Life (ASYM: 81.6 ± 16.1; SYM: 94.1 ± 9.7; P = .008). Knee flexion excursion was associated with knee functional recovery on the KOOS-Pain and the KOOS–Quality of Life ( P = .033 and P = .012, respectively) at 2 years after RTS, after controlling for the quadriceps strength LSI and graft type. Conclusion: Young athletes after ACLR with asymmetries in knee kinematics at the time of RTS reported decreased self-reported function 2 years later; however, the clinical importance of these differences needs to be further understood.

Pulmonary arterial stiffness is an independent risk factor for mortality in pulmonary hypertension (PH) and plays a critical role in PH pathophysiology. Our laboratory has recently demonstrated arterial stiffening early in experimental PH, along with evidence for a mechanobiological feedback loop by which arterial stiffening promotes further cellular remodeling behaviors (Liu F, Haeger CM, Dieffenbach PB, Sicard D, Chrobak I, Coronata AM, Suárez Velandia MM, Vitali S, Colas RA, Norris PC, Marinković A, Liu X, Ma J, Rose CD, Lee SJ, Comhair SA, Erzurum SC, McDonald JD, Serhan CN, Walsh SR, Tschumperlin DJ, Fredenburgh LE. JCI Insight 1: e86987, 2016). Cyclooxygenase-2 (COX-2) and prostaglandin signaling have been implicated in stiffness-mediated regulation, with prostaglandin activity inversely correlated to matrix stiffness and remodeling behaviors in vitro, as well as to disease progression in rodent PH models. The mechanism by which mechanical signaling translates to reduced COX-2 activity in pulmonary vascular cells is unknown. The present work investigated the transcriptional regulators Yes-associated protein (YAP) and WW domain-containing transcription regulator 1 (WWTR1, a.k.a., TAZ), which are known drivers of downstream mechanical signaling, in mediating stiffness-induced changes in COX-2 and prostaglandin activity in pulmonary artery smooth muscle cells (PASMCs). We found that YAP/TAZ activity is increased in PAH PASMCs and experimental PH and is necessary for the development of stiffness-dependent remodeling phenotypes. Knockdown of YAP and TAZ markedly induces COX-2 expression and downstream prostaglandin production by approximately threefold, whereas overexpression of YAP or TAZ reduces COX-2 expression and prostaglandin production to near undetectable levels. Together, our findings demonstrate a stiffness-dependent YAP/TAZ-mediated positive feedback loop that drives remodeling phenotypes in PASMCs via reduced COX-2 and prostaglandin activity. The ability to interrupt this critical mechanobiological feedback loop and enhance local prostaglandin activity via manipulation of YAP/TAZ signaling presents a highly attractive novel strategy for the treatment of PH.

Abstract FAM46C is one of the most recurrently mutated genes in multiple myeloma; however its role in disease pathogenesis has not been determined. Here we demonstrate that wild-type (WT) FAM46C overexpression induces substantial cytotoxicity in multiple myeloma cells. In contrast, FAM46C mutations found in multiple myeloma patients abrogate this cytotoxicity, indicating a survival advantage conferred by the FAM46C mutant phenotype. WT FAM46C overexpression downregulated IRF4, CEBPB, and MYC and upregulated immunoglobulin (Ig) light chain and HSPA5/BIP. Furthermore, pathway analysis suggests that enforced FAM46C expression activated the unfolded protein response pathway and induced mitochondrial dysfunction. CRISPR-mediated depletion of endogenous FAM46C enhanced multiple myeloma cell growth, decreased Ig light chain and HSPA5/BIP expression, activated ERK and antiapoptotic signaling, and conferred relative resistance to dexamethasone and lenalidomide treatments. Genes altered in FAM46C-depleted cells were enriched for signaling pathways regulating estrogen, glucocorticoid, B-cell receptor signaling, and ATM signaling. Together these results implicate FAM46C in myeloma cell growth and survival and identify FAM46C mutation as a contributor to myeloma pathogenesis and disease progression via perturbation in plasma cell differentiation and endoplasmic reticulum homeostasis. Cancer Res; 77(16); 4317–27. ©2017 AACR.