Mutation and cell state compatibility is required and targetable in Ph+ acute lymphoblastic leukemia minimal residual disease.

Efforts to cure BCR::ABL1 B cell acute lymphoblastic leukemia (Ph+ ALL) solely through inhibition of ABL1 kinase activity have thus far been insufficient despite the availability of tyrosine kinase inhibitors (TKIs) with broad activity against resistance mutants. The mechanisms that drive persistence within minimal residual disease (MRD) remain poorly understood and therefore untargeted. Utilizing 13 patient-derived xenograft (PDX) models and clinical trial specimens of Ph+ ALL, we examined how genetic and transcriptional features co-evolve to drive progression during prolonged TKI response. Our work reveals a landscape of cooperative mutational and transcriptional escape mechanisms that differ from those causing resistance to first generation TKIs. By analyzing MRD during remission, we show that the same resistance mutation can either increase or decrease cellular fitness depending on transcriptional state. We further demonstrate that directly targeting transcriptional state-associated vulnerabilities at MRD can overcome BCR::ABL1 independence, suggesting a new paradigm for rationally eradicating MRD prior to relapse. Finally, we illustrate how cell mass measurements of leukemia cells can be used to rapidly monitor dominant transcriptional features of Ph+ ALL to help rationally guide therapeutic selection from low-input samples.

In vivo bone marrow microenvironment siRNA delivery using lipid-polymer nanoparticles for multiple myeloma therapy.

Multiple myeloma (MM), a hematologic malignancy that preferentially colonizes the bone marrow, remains incurable with a survival rate of 3 to 6 mo for those with advanced disease despite great efforts to develop effective therapies. Thus, there is an urgent clinical need for innovative and more effective MM therapeutics. Insights suggest that endothelial cells within the bone marrow microenvironment play a critical role. Specifically, cyclophilin A (CyPA), a homing factor secreted by bone marrow endothelial cells (BMECs), is critical to MM homing, progression, survival, and chemotherapeutic resistance. Thus, inhibition of CyPA provides a potential strategy to simultaneously inhibit MM progression and sensitize MM to chemotherapeutics, improving therapeutic response. However, inhibiting factors from the bone marrow endothelium remains challenging due to delivery barriers. Here, we utilize both RNA interference (RNAi) and lipid-polymer nanoparticles to engineer a potential MM therapy, which targets CyPA within blood vessels of the bone marrow. We used combinatorial chemistry and high-throughput in vivo screening methods to engineer a nanoparticle platform for small interfering RNA (siRNA) delivery to bone marrow endothelium. We demonstrate that our strategy inhibits CyPA in BMECs, preventing MM cell extravasation in vitro. Finally, we show that siRNA-based silencing of CyPA in a murine xenograft model of MM, either alone or in combination with the Food and Drug Administration (FDA)-approved MM therapeutic bortezomib, reduces tumor burden and extends survival. This nanoparticle platform may provide a broadly enabling technology to deliver nucleic acid therapeutics to other malignancies that home to bone marrow.

A novel ß-catenin/BCL9 complex inhibitor blocks oncogenic Wnt signaling and disrupts cholesterol homeostasis in colorectal cancer.

Dysregulated Wnt/ß-catenin signaling is implicated in the pathogenesis of many human cancers, including colorectal cancer (CRC), making it an attractive clinical target. With the aim of inhibiting oncogenic Wnt activity, we developed a high-throughput screening AlphaScreen assay to identify selective small-molecule inhibitors of the interaction between ß-catenin and its coactivator BCL9. We identified a compound that consistently bound to ß-catenin and specifically inhibited in vivo native ß-catenin/BCL9 complex formation in CRC cell lines. This compound inhibited Wnt activity, down-regulated expression of the Wnt/ß-catenin signature in gene expression studies, disrupted cholesterol homeostasis, and significantly reduced the proliferation of CRC cell lines and tumor growth in a xenograft mouse model of CRC. This study has therefore identified a specific small-molecule inhibitor of oncogenic Wnt signaling, which may have value as a probe for functional studies and has important implications for the development of novel therapies in patients with CRC.

The Relationship Between Cardiorespiratory and Accelerometer-Derived Measures in Trail Running and the Influence of Sensor Location.

PURPOSE: To examine the relationship between cardiorespiratory and accelerometer-derived measures of exercise during trail running and determine the influence of accelerometer location. METHODS: Eight trail runners (7 males and 1 female; age 26 [5] y; maximal oxygen consumption [V˙O2] 70 [6] mL·kg-1·min-1) completed a 7-km trail run (elevation gain: 486 m), with concurrent measurements of V˙O2, heart rate, and accelerations recorded from 3 triaxial accelerometers attached at the upper spine, lower spine, and pelvis. External exercise intensity was quantified from the accelerometers using PlayerLoad™ per minute and accelerometry-derived average net force. External exercise volume was calculated using accumulated PlayerLoad and the product of average net force and duration (impulse). Internal intensity was calculated using heart rate and V˙O2-metrics; internal volume was calculated from total energy expenditure (work). All metrics were analyzed during both uphill (UH) and downhill (DH) sections of the trail run. RESULTS: PlayerLoad and average net force were greater during DH compared with UH for all sensor locations (P ≤ .004). For all accelerometer metrics, there was a sensor position × gradient interaction (F2,1429.003; P <.001). The upper spine was lower compared with both pelvis (P ≤ .003) and lower spine (P ≤ .002) for all accelerometer metrics during both UH and DH running. Relationships between accelerometer and cardiorespiratory measures during UH running ranged from moderate negative to moderate positive (r = -.31 to .41). Relationships were stronger during DH running where there was a nearly perfect correlation between work and impulse (r = .91; P < .001). CONCLUSIONS: Simultaneous monitoring of cardiorespiratory and accelerometer-derived measures during trail running is suggested because of the disparity between internal and external intensities during changes in gradient. Sensor positioning close to the center of mass is recommended.

miR-15a/16-1 deletion in activated B cells promotes plasma cell and mature B-cell neoplasms.

Chromosome 13q deletion [del(13q)], harboring the miR-15a/16-1 cluster, is one of the most common genetic alterations in mature B-cell malignancies, which originate from germinal center (GC) and post-GC B cells. Moreover, miR-15a/16 expression is frequently reduced in lymphoma and multiple myeloma (MM) cells without del(13q), suggesting important tumor-suppressor activity. However, the role of miR-15a/16-1 in B-cell activation and initiation of mature B-cell neoplasms remains to be determined. We show that conditional deletion of the miR-15a/16-1 cluster in murine GC B cells induces moderate but widespread molecular and functional changes including an increased number of GC B cells, percentage of dark zone B cells, and maturation into plasma cells. With time, this leads to development of mature B-cell neoplasms resembling human extramedullary plasmacytoma (EP) as well as follicular and diffuse large B-cell lymphomas. The indolent nature and lack of bone marrow involvement of EP in our murine model resembles human primary EP rather than MM that has progressed to extramedullary disease. We corroborate human primary EP having low levels of miR-15a/16 expression, with del(13q) being the most common genetic loss. Additionally, we show that, although the mutational profile of human EP is similar to MM, there are some exceptions such as the low frequency of hyperdiploidy in EP, which could account for different disease presentation. Taken together, our studies highlight the significant role of the miR-15a/16-1 cluster in the regulation of the GC reaction and its fundamental context-dependent tumor-suppression function in plasma cell and B-cell malignancies.

BCL9 provides multi-cellular communication properties in colorectal cancer by interacting with paraspeckle proteins.

Colorectal cancer (CRC) is the third most commonly diagnosed cancer, which despite recent advances in treatment, remains incurable due to molecular heterogeneity of tumor cells. The B-cell lymphoma 9 (BCL9) oncogene functions as a transcriptional co-activator of the Wnt/ß-catenin pathway, which plays critical roles in CRC pathogenesis. Here we have identified a ß-catenin-independent function of BCL9 in a poor-prognosis subtype of CRC tumors characterized by expression of stromal and neural associated genes. In response to spontaneous calcium transients or cellular stress, BCL9 is recruited adjacent to the interchromosomal regions, where it stabilizes the mRNA of calcium signaling and neural associated genes by interacting with paraspeckle proteins. BCL9 subsequently promotes tumor progression and remodeling of the tumor microenvironment (TME) by sustaining the calcium transients and neurotransmitter-dependent communication among CRC cells. These data provide additional insights into the role of BCL9 in tumor pathogenesis and point towards additional avenues for therapeutic intervention.

Orbital Volume Correction in Orbital Floor Fractures: A Comparison of Transorbital and Transantral Techniques.

PURPOSE: We compared the accuracy of orbital volume correction between the transorbital and transantral reconstructive techniques. MATERIALS AND METHODS: A retrospective cohort study was performed of patients who had undergone repair of isolated, unilateral orbital floor blowout fractures at Legacy Emanuel Hospital from 2013 to 2018. A total of 21 patients were identified and included in the predictor variable cohorts of the transorbital versus transantral repair technique. The outcome variable of orbital volume correction was evaluated by comparing the volume of the postoperative repaired orbits with that of the contralateral noninjured orbits. Additional ordinal variables analyzed included the preoperative orbital defect size and analysis of the transantral cohort stratified by the plating technique used. Data were assessed using analysis of variance and paired t tests. RESULTS: A transantral approach was used for orbital repair in 9 patients. In these patients, the postoperative orbital volume in the injured orbit was 2.69% greater than that in the uninjured orbit. The 12 patients who had undergone transantral repair had a postoperative orbital volume in the injured orbit that was 0.56% smaller than that of the uninjured orbit (P = .033). Division of the transantral cohort into 2 different plating techniques identified a less than 1% difference in mean orbital volume correction between the 2 techniques (P = .104). The average defect volume before transorbital repair was 4.87 cm3 compared with 5.22 cm3 for transantral repair (P = .907). CONCLUSIONS: The results from the present study have shown that the accuracy of orbital volume correction using the transantral approach will be comparable to that of the transorbital approach, as shown by a small, but statistically significant, increased accuracy in the volume correction with the transantral approach. Additional investigation to establish clinical correlations with these findings should be conducted.

Human MYD88L265P is insufficient by itself to drive neoplastic transformation in mature mouse B cells.

MYD88 L265P is the most common mutation in lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (LPL/WM) and one of the most frequent in poor-prognosis subtypes of diffuse large B-cell lymphoma (DLBCL). Although inhibition of the mutated MYD88 pathway has an adverse impact on LPL/WM and DLBCL cell survival, its role in lymphoma initiation remains to be clarified. We show that in mice, human MYD88L265P promotes development of a non-clonal, low-grade B-cell lymphoproliferative disorder with several clinicopathologic features that resemble human LPL/WM, including expansion of lymphoplasmacytoid cells, increased serum immunoglobulin M (IgM) concentration, rouleaux formation, increased number of mast cells in the bone marrow, and proinflammatory signaling that progresses sporadically to clonal, high-grade DLBCL. Murine findings regarding differences in the pattern of MYD88 staining and immune infiltrates in the bone marrows of MYD88 wild-type (MYD88WT) and MYD88L265P mice are recapitulated in the human setting, which provides insight into LPL/WM pathogenesis. Furthermore, histologic transformation to DLBCL is associated with acquisition of secondary genetic lesions frequently seen in de novo human DLBCL as well as LPL/WM-transformed cases. These findings indicate that, although the MYD88L265P mutation might be indispensable for the LPL/WM phenotype, it is insufficient by itself to drive malignant transformation in B cells and relies on other, potentially targetable cooperating genetic events for full development of lymphoma.

How well do we manage the odontogenic keratocyst? A multicenter study.

OBJECTIVE: The aim of this study was to answer the following clinical questions: Among patients treated for odontogenic keratocysts (OKCs), what is the overall 5-year disease-free rate, and what factors are associated with disease recurrence? STUDY DESIGN: We implemented a multicenter retrospective cohort study composed of patients presenting for the evaluation and management of previously untreated OKCs. The predictor variables were grouped into demographic, medical, radiographic, and operative categories. The primary outcome variable was time to lesion recurrence. Data analyses were performed by using bivariate analysis and univariate or multivariate Cox proportional hazards models. RESULTS: The study sample was composed of 231 OKCs. Of these, 57 (24.7%) were treated with decompression with residual cystectomy, 86 (37.2%) with enucleation without adjuvant therapy, and 78 (33.8%) with enucleation with peripheral ostectomy. There were 44 recurrences (19%), with a median time to recurrence of 26.7 months (range 15.8-49.8). CONCLUSIONS: This multicenter study is the largest study analyzing disease recurrence after treatment of OKCs by using appropriate statistical analysis for a time-to-event outcome (disease recurrence). The 5-year disease-free estimate was 29%. Mandibular lesions, multilocular lesions, and lesions treated with decompression and residual cystectomy were associated with recurrence.

Constitutive Ras signaling and Ink4a/Arf inactivation cooperate during the development of B-ALL in mice.

Despite recent advances in treatment, human precursor B-cell acute lymphoblastic leukemia (B-ALL) remains a challenging clinical entity. Recent genome-wide studies have uncovered frequent genetic alterations involving RAS pathway mutations and loss of the INK4A/ARF locus, suggesting their important role in the pathogenesis, relapse, and chemotherapy resistance of B-ALL. To better understand the oncogenic mechanisms by which these alterations might promote B-ALL and to develop an in vivo preclinical model of relapsed B-ALL, we engineered mouse strains with induced somatic KrasG12D pathway activation and/or loss of Ink4a/Arf during early stages of B-cell development. Although constitutive activation of KrasG12D in B cells induced prominent transcriptional changes that resulted in enhanced proliferation, it was not sufficient by itself to induce development of a high-grade leukemia/lymphoma. Instead, in 40% of mice, these engineered mutations promoted development of a clonal low-grade lymphoproliferative disorder resembling human extranodal marginal-zone lymphoma of mucosa-associated lymphoid tissue or lymphoplasmacytic lymphoma. Interestingly, loss of the Ink4a/Arf locus, apart from reducing the number of apoptotic B cells broadly attenuated KrasG12D-induced transcriptional signatures. However, combined Kras activation and Ink4a/Arf inactivation cooperated functionally to induce a fully penetrant, highly aggressive B-ALL phenotype resembling high-risk subtypes of human B-ALL such as BCR-ABL and CRFL2-rearranged. Ninety percent of examined murine B-ALL tumors showed loss of the wild-type Ink4a/Arf locus without acquisition of highly recurrent cooperating events, underscoring the role of Ink4a/Arf in restraining Kras-driven oncogenesis in the lymphoid compartment. These data highlight the importance of functional cooperation between mutated Kras and Ink4a/Arf loss on B-ALL.