Publications

2019
Rinaldo L, Brown DA, Bhargav AG, Rusheen AE, Naylor RM, Gilder HE, Monie DD, Youssef SJ, Parney IF. Venous thromboembolic events in patients undergoing craniotomy for tumor resection: incidence, predictors, and review of literature. J Neurosurg. 2019 :1-12.Abstract
OBJECTIVEThe authors sought to investigate the incidence and predictors of venous thromboembolic events (VTEs) after craniotomy for tumor resection, which are not well established, and the efficacy of and risks associated with VTE chemoprophylaxis, which remains controversial.METHODSThe authors investigated the incidence of VTEs in a consecutive series of patients presenting to the authors' institution for resection of an intracranial lesion between 2012 and 2017. Information on patient and tumor characteristics was collected and independent predictors of VTEs were determined using stepwise multivariate logistic regression analysis. Review of the literature was performed by searching MEDLINE using the keywords "venous thromboembolism," "deep venous thrombosis," "pulmonary embolism," "craniotomy," and "brain neoplasms."RESULTSThere were 1622 patients included for analysis. A small majority of patients were female (52.6%) and the mean age of the cohort was 52.9 years (SD 15.8 years). A majority of intracranial lesions were intraaxial (59.3%). The incidence of VTEs was 3.0% and the rates of deep venous thromboses and pulmonary emboli were 2.3% and 0.9%, respectively. On multivariate analysis, increasing patient age (unit OR 1.02, 95% CI 1.00-1.05; p = 0.018), history of VTE (OR 7.26, 95% CI 3.24-16.27; p < 0.001), presence of motor deficit (OR 2.64, 95% CI 1.43-4.88; p = 0.002), postoperative intracranial hemorrhage (OR 4.35, 95% CI 1.51-12.55; p < 0.001), and prolonged intubation or reintubation (OR 3.27, 95% CI 1.28-8.32; p < 0.001) were independently associated with increased odds of a VTE. There were 192 patients who received VTE chemoprophylaxis (11.8%); the mean postoperative day of chemoprophylaxis initiation was 4.6 (SD 3.8). The incidence of VTEs was higher in patients receiving chemoprophylaxis than in patients not receiving chemoprophylaxis (8.3% vs 2.2%; p < 0.001). There were 30 instances of clinically significant postoperative hemorrhage (1.9%), with only 1 hemorrhage occurring after initiation of VTE chemoprophylaxis (0.1%).CONCLUSIONSThe study results show the incidence and predictors of VTEs after craniotomy for tumor resection in this patient population. The incidence of VTE within this cohort appears low and comparable to that observed in other institutional series, despite the lack of routine prophylactic anticoagulation in the postoperative setting.
2017
Monie DD, DeLoughery EP. Pathogenesis of thrombosis: cellular and pharmacogenetic contributions. Cardiovasc Diagn Ther [Internet]. 2017;7 (Suppl 3) :S291-S298. Publisher's VersionAbstract
Our understanding of thrombosis formation has evolved significantly ever since physician Rudolf Virchow proposed his “triad” theory in 1856. Modern science has elucidated the mechanisms of stasis, hypercoagulability, and endothelial dysfunction. Today, we have a firm understanding of the key molecular factors involved in the coagulation cascade and fibrinolytic system, as well as the underlying genetic influences. This knowledge of cellular and genetic contributors has been translated into diverse pharmaceutical interventions. Here, we examine the molecular and cellular mechanisms of thrombosis and its associated pathologies. We also review the current state of pharmacologic interventions, including pro- and anti-thrombotics, direct oral anticoagulants, and anti-platelet therapies. The pharmacogenetic factors that guide clinical decision making and prognosis are described in detail. Finally, we explore new approaches to thrombosis drug discovery, repurposing, and diagnostics. We argue that network biology tools will enable a systems pharmacology revolution in the next generation of interventions, facilitating precision medicine applications and ultimately leading to improved patient outcomes.
2015
Monie DD, Bhatia SK. Bioprinting of Dynamic Human Organs-on-Chips: Enabling Technologies for Rapid Drug Development and Personalized Medicine. In: Turksen K Bioprinting in Regenerative Medicine. 1st ed. New York: Springer ; 2015. pp. 123-137. Publisher's VersionAbstract

Dynamic human organs-on-chips are microfluidic devices capable of reproducing the physiology of human organs, accurately simulating both normal and disease states. These microphysiological systems offer more predictive and ethical alternatives both to animal-based preclinical drug studies and to cell-based in vitro diagnostics. But multiple organs-on-chips must be connected together in order to emulate the full human body. Therefore, new and more complex organs-on-chips that are more representative and versatile need to be designed and developed. The most salient challenge lies in their fabrication. Current prototypes rely on traditional microfluidic and microelectromechanical system methods that are too time-consuming for rapid prototyping, troubleshooting, and customization. Newer three-dimensional bioprinting technologies can help facilitate fabrication and deployment of next generation organs-on-chips. This review of the literature provides background on the clinical utility of dynamic human organs-on-chips. It lays out their design and fabrication requirements in the context of state of the art bioprinting technologies. It also proposes development pathways for bioprinted organs-on-chips to engineer more predictive designs. Finally, regulatory considerations and the impact of future bioprinting approaches on organ-on-chips are discussed with emphasis on enabling an era of personalized medicine.

2012
Impellizzeri D, Mazzon E, Paola DR, Galuppo M, Bramanti P, Zhang J, Bobb K, Monie D, Meshulam J, Sliskovic DR, et al. PBS-1086, a Rel inhibitor of NF-κB, ameliorates collagen-induced arthritis in mice. European Journal of Inflammation. 2012;10 (1) :51-59.Abstract

The family of nuclear factor-kappaB (NF-κB) transcription factors is intimately involved in the regulation of expression of numerous genes in the setting of the inflammatory response. Inflammation, cartilage degradation, cell proliferation, angiogenesis and pannus formation are hallmarks of the pathogenesis of both collagen-induced arthritis (CIA) in rodents and rheumatoid arthritis (RA) in humans. The aim of this study is to investigate the effect of PBS-1086, a Rel inhibitor of NF-κB, on the modulation of the inflammatory response in mice subjected to CIA in comparison to the effect of etanercept. CIA was induced in mice by an intradermal injection of bovine type II collagen (CII) emulsion and complete Freund’s adjuvant (CFA) at the base of the tail. On day 21, a second injection of CII in CFA was administered. Mice developed erosive hind paw arthritis when immunised with CII in CFA. Macroscopic clinical evidence of CIA first appeared as peri-articular erythema and oedema in the hind paws. The incidence of CIA was 100% by day 28 in the CII challenged mice and the severity of CIA progressed over a 35-day period with a resorption of bone. The histopathology of CIA included erosion of the cartilage at the joint. Treatment with PBS-1086 starting at the onset of arthritis (day 21) ameliorated the clinical signs at days 21-35 and improved histological status in the joint and paw. In addition, it also reduced the neutrophil infiltration which is a key mediator of RA. In this study, we demonstrate that PBS-1086 exerts an anti-inflammatory effect during chronic inflammation and ameliorates the tissue damage associated with CIA. The anti-inflammatory activities of PBS-1086 are comparable to those of etanercept treatment.

2011
Oh U, McCormick MJ, Datta D, Turner RV, Bobb K, Monie DD, Sliskovic RD, Tanaka Y, Zhang J, Meshulam J, et al. Inhibition of immune activation by a novel nuclear factor-kappa B inhibitor in HTLV-I-associated neurologic disease. Blood. 2011;117 (12) :3363-9.Abstract
The human T-lymphotropic virus type I (HTLV-I) causes a chronic inflammatory disorder of the central nervous system termed HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). HTLV-I encodes a protein known to activate several host-signaling pathways involved in inflammation, such as the nuclear factor-κB (NF-κB). The contribution of the NF-κB pathway to the pathogenesis of HAM/TSP, however, has not been fully defined. We show evidence of canonical NF-κB activation in short-term cultures of peripheral blood mononuclear cells (PBMCs) from subjects with HAM/TSP. NF-κB activation was closely linked to HTLV-I viral protein expression. The NF-κB activation in HAM/TSP PBMCs was reversed by a novel small-molecule inhibitor that demonstrates potent and selective NF-κB antagonist activity. Inhibition of NF-κB activation led to a reduction in the expression of lymphocyte activation markers and resulted in reduced cytokine signaling in HAM/TSP PBMCs. Furthermore, NF-κB inhibition led to a reduction in spontaneous lymphoproliferation, a key ex vivo correlate of the immune activation associated with HAM/TSP. These results indicate that NF-κB activation plays a critical upstream role in the immune activation of HAM/TSP, and identify the NF-κB pathway as a potential target for immunomodulation in HAM/TSP.
McCormick M, Oh U, Datta D, Turner R, Bobb K, Monie D, Sliskovic D, Zhang J, Meshulam J, Jacobson S. NFkB activation promotes immune activation in HTLV-I-associated myelopathy / tropical spastic paraparesis. Retrovirology [Internet]. 2011;8 (Suppl 1) :A117. Publisher's VersionAbstract

Evidence suggests that HTLV-I-induced immune activation plays a key role in the pathogenesis of HAM/TSP. The HTLV-I-encoded transactivating protein Tax is known to activate nuclear factor kappa B (NFkB), a key host signaling pathway regulating immune response, but the contribution of the NFkB pathway to the immune activation associated with HAM/TSP has yet to be fully defined. We examined NFkB activation in peripheral blood mononuclear cells (PBMC) from subjects with HAM/TSP, and tested the effect of NFkB inhibition on key ex vivo correlates of immune activation in HAM/TSP.

We examined the role of NFkB activation during immune activation associated with HAM/TSP by using small molecule NFkB inhibitors, including a newly developed selective inhibitor of NFkB, PBS-1086.

NFkB activation was assessed in peripheral-blood mononuclear cells (PBMC) from subjects with HAM/TSP and in healthy donors (HD). Nuclear translocation of the NFkB RelA was significantly higher in PBMC from subjects with HAM/TSP compared to HD (p=0.032) following short-term (20 h) culture, indicating increased activation of the NFkB pathway in HAM/TSP. Treatment with the small molecule inhibitor PBS-1086 reduced NFkB activation (p<0.01). PBS-1086 reduced expression of CD25 and CD69 in HAM/TSP PBMC as well as phosphorylation of STAT5 in a dose-dependent manner (p<0.01 for all). PBS-1086 also inhibited spontaneous lymphoproliferation of HAM/TSP PBMC in a dose-dependent manner (p=0.0286). PBS-1086 treatment resulted in a mean proviral load reduction of 20% compared to untreated PBMC in a 72 h culture.

These results indicate that NFkB activation plays a critical upstream role in the immune activation associated with HAM/TSP, and identify the NFkB pathway as a potential therapeutic target for immune modulation in HAM/TSP.

2010
Oh U, McCormick M, Datta D, Turner R, Bobb K, Monie D, Sliskovic RD, Zhang J, Meshulam J, Jacobson S. NFkB activation promotes immune activation in HTLV-I-associated myelopathy/tropical spastic paraparesis. Journal of NeuroVirology [Internet]. 2010;16 (Suppl 1) :63–64. Publisher's VersionAbstract

Background: Human T lymphotropic virus type I (HTLV-I) is the etiologic agent of HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), a chronic inflammatory disorder of the central nervous system. Evidence suggests that viral-induced immune activation plays a key role in the pathogenesis of HAM/TSP. The HTLV-I-encoded transactivating protein Tax is known to activate nuclear factor kappa B (NFkB), a key host signaling pathway regulating immune response, but the contribution of the NFkB pathway to the immune activation associated with HAM/TSP has yet to be fully defined. To further delineate the role of NFkB activation in HAM/TSP, we examined NFkB activation in peripheral-blood mononuclear cells (PBMC) from subjects with HAM/TSP, and tested the effect of NFkB inhibition on key ex vivo correlates of immune activation in HAM/TSP.

Materials and Methods: We examined the role of NFkB activation during immune activation associated with HAM/TSP by using small molecule NFkB inhibitors, including a newly developed selective inhibitor of NFkB, PBS-1086. NFkB activation was measured by a DNA-binding ELISA on nuclear extracts from PBMC of subjects with HAM/TSP and healthy donors to detect nuclear translocation of NFkB subunits. Immune activation in HAM/TSP PBMC was measured by tritiated thymidine incorporation, a marker for spontaneous proliferation, and by FACS analysis of the lymphocyte activation markers CD25 and CD69 and the pro-inflammatory cytokine STAT5. Proviral loads in untreated and inhibitor-treated HAM/TSP samples were measured by real time PCR.

Results: NFkB activation was assessed in peripheral-blood mononuclear cells (PBMC) from subjects with HAM/TSP and in healthy donors (HD). Nuclear translocation of the NFkB RelA was significantly higher in PBMC from subjects with HAM/TSP compared to HD (p = 0.032) following short-term (20 h) culture, indicating increased activation of the NFkB pathway in HAM/TSP. Treatment with the small molecule inhibitor PBS-1086 reduced NFkB activation (p < 0.01). PBS-1086 reduced expression of CD25 and CD69 in HAM/TSP PBMC as well as phosphorylation of STAT5 in a dose-dependent manner (p < 0.01 for all). PBS-1086 also inhibited spontaneous lymphoproliferation of HAM/TSP PBMC in a dose-dependent manner. PBS-1086treatment resulted in a mean proviral load reduction of 20% compared to untreated PBMC in a 72 h culture.

Discussion: These results indicate that NFkB activation plays a critical upstream role in the immune activation associated with HAM/TSP, and identify the NFkB pathway as a potential therapeutic target for immune modulation in HAM/TSP.

Hunter JE, Willmore E, Irving JAE, Sliskovic RD, Monie D, Bobb K, Zhang J, Durkacz BW. Abstract 3561: The radiosensitizing and cytotoxic effects of PBS-1086, a Rel inhibitor of NF-KB, in breast cancer cells. Cancer Research [Internet]. 2010;70 :3561. Publisher's VersionAbstract
Recent RNA interference screening has identified p53 and ras mutations to be synthetic lethal with the canonical and non-canonical pathways of NF-κB. We have synthesized inhibitors that antagonize bindings of RelA (p65), RelB and c-rel to the NF-κB sites and thus inhibit both the canonical and non-canonical paths. One such compound, PBS-1086, is evaluated here as a radiosensitizer in wild type and p65−/− mouse embryonic fibroblasts (MEFs), and as a radiosensitizing and chemotherapeutic agent in the p53 deficient and KRAS (G13D) mutant breast cancer cell line, MDA-MB-231.We found that p65−/− MEFs were ≥ 2-fold more sensitive than p65+/+ MEFs to irradiation (IR) in clonogenic survival assays, thus demonstrating that NF-κB activation conferred radio-resistance. Incubation with PBS-1086 at 1.4 µM, a non-cytotoxic concentration, radiosensitized p65+/+ MEFs (LD50 value of 1.5 Gy for IR + PBS-1086 compared with 2.3 Gy for IR alone). PBS-1086 had no effect on radio-sensitization of p65−/− MEFs, which indicated the radiosensitizing effect of PBS-1086 was mediated through p65. Similar radiosensitizing effects were observed by using RNA interference technique to down-regulate the expression level of p65 by > 95% in p65+/+ MEFs but not in p65−/− MEFs. We also confirmed the inhibition of the NF-κB by PBS-1086 or p65 siRNA in MEFs. NF-κB-dependent luciferase expression increased by 2.5 fold after 10 Gy IR in p65+/+ MEFs. Treatment with PBS-1086 or p65 siRNA reduced luciferase expression significantly. Similarly, IR increased p65+/+ MEFs nuclear extract NF-κB DNA binding activities by 2 fold, which was significantly decreased by either PBS-1086 incubation or p65 siRNA treatment in these cells.Next, we studied PBS-1086 in MDA-MB-231 breast cancer cells with constitutive NF-κB activation. First, we tested PBS-1086 as a single agent on the growth of the breast cancer cells. Significant cytotoxic effect to PBS-1086 was observed with an LD50 value of 0.4 μM to the MDA-MB-231 cells. Then we tested synergistic effect of PBS-1086 with irradiation in these cells. Incubation with 0.2 µM PBS-1086 significantly enhanced radiation effect by 1.5 fold (LD50 value of 1.66 Gy IR for IR + PBS-1086 compare with 2.55 Gy for IR alone).In conclusion, we have demonstrated that a Rel inhibitor of NF-KB can potentiate radioactivity through blocking the canonical pathway. It can also selectively kill tumor cells with p53 and KRAS mutations, perhaps through blocking the canonical and non-canonical pathways.Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3561.
2009
Kelly JR, Rubin AJ, Davis JH, Ajo-Franklin CM, Cumbers J, Czar MJ, de Mora K, Glieberman AL, Monie DD, Endy D. Measuring the activity of BioBrick promoters using an in vivo reference standard. J Biol Eng. 2009;3 :4.Abstract
BACKGROUND: The engineering of many-component, synthetic biological systems is being made easier by the development of collections of reusable, standard biological parts. However, the complexity of biology makes it difficult to predict the extent to which such efforts will succeed. As a first practical example, the Registry of Standard Biological Parts started at MIT now maintains and distributes thousands of BioBrick standard biological parts. However, BioBrick parts are only standardized in terms of how individual parts are physically assembled into multi-component systems, and most parts remain uncharacterized. Standardized tools, techniques, and units of measurement are needed to facilitate the characterization and reuse of parts by independent researchers across many laboratories. RESULTS: We found that the absolute activity of BioBrick promoters varies across experimental conditions and measurement instruments. We choose one promoter (BBa_J23101) to serve as an in vivo reference standard for promoter activity. We demonstrated that, by measuring the activity of promoters relative to BBa_J23101, we could reduce variation in reported promoter activity due to differences in test conditions and measurement instruments by approximately 50%. We defined a Relative Promoter Unit (RPU) in order to report promoter characterization data in compatible units and developed a measurement kit so that researchers might more easily adopt RPU as a standard unit for reporting promoter activity. We distributed a set of test promoters to multiple labs and found good agreement in the reported relative activities of promoters so measured. We also characterized the relative activities of a reference collection of BioBrick promoters in order to further support adoption of RPU-based measurement standards. CONCLUSION: Relative activity measurements based on an in vivoreference standard enables improved measurement of promoter activity given variation in measurement conditions and instruments. These improvements are sufficient to begin to support the measurement of promoter activities across many laboratories. Additional in vivo reference standards for other types of biological functions would seem likely to have similar utility, and could thus improve research on the design, production, and reuse of standard biological parts.
2008
Ducker C, Fouts TR, Monie DD. Compositions and methods for treating viral infections. [Internet]. 2008. Publisher's Version
2007
Merbs SL, Hackler, L. J, Khan MA, Monie DD, Zack DJ. Expression of Photoreceptor-Specific Genes Correlates With DNA Hypomethylation. Invest. Ophthalmol. Vis. Sci. [Internet]. 2007;48 :3778. Publisher's VersionAbstract

Purpose: One element of transcriptional regulation is the local epigenetic configuration of a gene. DNA methylation is an epigenetic phenomenon that controls various genomic functions without altering the DNA sequence. The promoter sequences of housekeeping genes are typically unmethylated, and aberrant methylation is known to shut down expression of tumor suppressor genes in cancer. To investigate whether DNA methylation might play a role in tissue-specific expression of photoreceptor genes, we investigated the methylation status of photoreceptor-specific genes and correlated the degree of methylation with the expression of those genes.

Methods: Genomic DNA from WERI and Y79 human retinoblastoma cell lines and 293 cells were treated with bisulfite. Primers were designed to amplify only the bisulfite modified DNA in 300-500bp segments of the 1000bp upstream of the TSS through the first exon of selected photoreceptor-specific genes (RPB3, RHO, OPN1SW, OPN1MW, and OPN1LW). Direct sequencing of the PCR products was performed. The ratio of unmethylated/methylated (T/C) DNA was calculated for each CpG site. Expression of the same genes was measured by real time PCR. The cell line experiments were then reproduced in the mouse, comparing the methylation status of RBP3, RHO, OPN1SW and OPN1MW in retina, brain, kidney and testes with the expression level of those genes.

Results: None of the photoreceptor-specific genes were detected in 293 cells, Y79 cells expressed RPB3 but only very low levels of the opsins, and WERI cells expressed RBP3, OPN1MW and OPN1LW. The CpG sites 1000bp upstream of the TSS through the end of the first exon in all photoreceptor-specific genes were primarily methylated in 293 cells, consistent with the non-expression. In Y79 cells, the photoreceptor-specific genes were primarily methylated, except for RPB3, which was primarily unmethylated. The RPB3, OPN1MW, and OPN1LW genes were primarily unmethylated in WERI cells, and the RHO and OPN1SW genes were methylated. The same correlation between expression and the lack of methylation was found in corresponding experiments in mice. Photoreceptor-specific genes were overall less methylated in the retina, than in the other non-expressing tissues.

Conclusions: We have demonstrated that the relative lack of DNA methylation in some photoreceptor-specific genes correlates with the expression of those genes, both in human tissue culture cells and in several mouse tissues. Whether the methylation in part directs the tissue-specific non-expression, or is simply a consequence of the non-expression is yet to be determined.

Monie D. Pulmonary Hypertension. In: Prasad R, Kahan S In a Page Cardiology. Philadelphia: Lippincott Williams & Wilkins ; 2007. pp. 148-149.
1999
Vath GM, Earhart CA, Monie DD, Iandolo JJ, Schlievert PM, Ohlendorf DH. The crystal structure of exfoliative toxin B: a superantigen with enzymatic activity. Biochemistry. 1999;38 (32) :10239-46.Abstract
The exfoliative toxins (ETs) cause staphylococcal scalded skin syndrome, a disease characterized by specific separation of layers of the skin. Evidence suggests that the toxins act as serine proteases, though the specific substrate and mode of action are not known for certain. The crystal structure of exfoliative toxin A (ETA) was reported earlier and shown to be similar to that of the chymotrypsin-like serine proteases. Here, we report the 2.4 A resolution crystal structure of the other exfoliative toxin, ETB, which is 40% identical to ETA. The overall structures of ETA and ETB are similar including the positions of key residues within the active site. The structure of ETB supports the previous findings that the ETs are serine proteases that cleave substrates after glutamic acid residues. In this study we also discuss a number of structural differences including a large 14 residue loop insertion which may be a key feature involved in the differing biological properties of the ETs, particularly the pyrogenic and lethal activities of ETB not shared by ETA.
Monday SR, Vath GM, Ferens WA, Deobald C, Rago JV, Gahr PJ, Monie DD, Iandolo JJ, Chapes SK, Davis WC, et al. Unique superantigen activity of staphylococcal exfoliative toxins. J Immunol. 1999;162 (8) :4550-9.Abstract
Certain strains of Staphylococcus aureus express one or both of two related, but immunologically distinct, exfoliative toxins (ETA and ETB). These toxins induce the symptoms associated with staphylococcal scalded skin syndrome. Both ETs have been shown to stimulate T cell proliferation. Recently, it was reported that ETA is a superantigen that stimulates T cells bearing human Vbeta2 or several murine Vbetas. However, other investigators have proposed that the superantigenicity reported for ETA resulted from contaminants in commercial preparations. This present study addresses those conflicting reports by assessing the biological and immunologic activities of highly purified rETs. ETA and ETB required APCs to induce selective polyclonal expansion of several human Vbetas (huVbetas), although, neither toxin expanded huVbeta2. ETB induced expansion of murine T cells bearing Vbetas 7 and 8, those that have the highest homology to the huVbetas expanded by ETA and ETB. Although flow cytometry of ETB-stimulated T cells matched PCR results, stimulation by ETA reduced percentages of T cells positive for several huVbetas that had been shown to have increased levels of mRNA transcripts. ETA and ETB induced contrasting reactions in vivo. In rabbits, ETB was moderately pyrogenic and enhanced susceptibility to lethal shock, while ETA lacked both activities. Predictions based on comparisons with other superantigens suggest molecular regions potentially involved in receptor binding in the ETA crystal structure and a modeled ETB three-dimensional structure. These results show that ETs are superantigens with unique properties that could account for the discrepancies reported.