Target Mucin 1 Research Articles

MUC1 Drives the Progression and Chemoresistance of Clear Cell Renal Carcinomas.

While the transmembrane glycoprotein mucin 1 (MUC1) is clustered at the apical borders of normal epithelial cells, with transformation and loss of polarity, MUC1 is found at high levels in the cytosol and is uniformly distributed over the entire surface of carcinoma cells, where it can promote tumor progression and adversely affects the response to therapy. Clear cell renal cell carcinoma (ccRCC), the main histotype of kidney cancer, is typically highly resistant to conventional and targeted therapies for reasons that remain largely unknown. In this context, we investigated whether MUC1 also plays a pivotal role in the cellular and molecular events driving ccRCC progression and chemoresistance. We showed, using loss- and gain-of-function approaches in ccRCC-derived cell lines, that MUC1 not only influences tumor progression but also induces a multi-drug-resistant profile reminiscent of the activation of ABC drug efflux transporters. Overall, our results suggest that targeting MUC1 may represent a novel therapeutic approach to limit ccRCC progression and improve drug sensitivity.

BLV-miR-B1-5p Promotes Staphylococcus aureus Adhesion to Mammary Epithelial Cells by Targeting MUC1.

Bovine leukemia virus (BLV) is widely prevalent worldwide and can persistently infect mammary epithelial cells in dairy cows, leading to reduced cellular antimicrobial capacity. BLV-encoded microRNAs (BLV-miRNAs) can modify host genes and promote BLV replication. We previously showed that BLV-miR-B1-5p significantly promoted Staphylococcus aureus (S. aureus) adhesion to bovine mammary epithelial (MAC-T) cells; however, the pathway responsible for this effect remained unclear. This study aims to examine how BLV-miR-B1-5p promotes S. aureus adhesion to MAC-T cells via miRNA target gene prediction and validation. Target site prediction showed that BLV-miR-B1-5p could target the mucin family gene mucin 1 (MUC1). Real-time polymerase chain reaction, immunofluorescence, and dual luciferase reporter assay further confirmed that BLV-miR-B1-5p could target and inhibit the expression of MUC1 in bovine MAC-T cells while interfering with the expression of MUC1 promoted S. aureus adhesion to MAC-T cells. These results indicate that BLV-miR-B1-5p promotes S. aureus adhesion to mammary epithelial cells by targeting MUC1.

Tandem CAR-T cells targeting MUC1 and PSCA combined with anti-PD-1 antibody exhibit potent preclinical activity against non-small cell lung cancer

Chimeric antigen receptor (CAR)-T cells encounter many issues when treating solid tumors, including tumor antigen heterogeneity and immunosuppression. United targeting of two tumor-associated antigens (TAAs) and blocking of PD-1 may solve this problem and enhance the function of CAR-T. Mucin 1 (MUC1) and prostate stem cell antigen (PSCA) are overexpressed in non-small cell lung cancer (NSCLC). Here, we constructed a bivalent tandem CAR-T (Tan CAR-T), which can simultaneously target MUC1 and PSCA and evaluated its effects of inhibiting non-small cell lung cancer (NSCLC) in vitro and in vivo. Results indicated that the tumor killing effect of these Tan CAR-T was more effective than that of single-target CAR-T, its antitumor efficacy could be further strengthened by anti-PD-1 antibody. Our study reported a previously unstudied therapeutic effect of a Tan CAR-T in NSCLC, providing a preclinical rationale for anti-PD-1 antibody combined with Tan CAR-T targeting MUC1 and PSCA in the treatment of NSCLC.

High-efficiency interparticle DNA walker amplifier combined with N-Ti3C2 quantum dots as electrochemiluminescence emitters for sensitive protein assay

In this work, an electrochemiluminescence (ECL) biosensor based on interparticle DNA walker as effective signal amplification strategy and N-Ti3C2 QDs as high-efficiency emitters was designed for sensitive assay of mucin 1 (MUC1), which is closely related to the occurrence and development of tumors. First, an interparticle DNA walker which contains walking particles (WPs) and orbit particles (OPs) was fabricated. In the presence of target MUC1 and DNAzyme, the WPs can walk along the surface of OPs, and they can continue to walk along next OPs after completing previous walk. As the moving freedom and area improved, the interparticle DNA walker could effectively improve the conversion efficiency of target. Furthermore, N-Ti3C2 QDs with high ECL luminous efficiency were synthesized by using polyethyleneimine (PEI) as nitrogen source. Due to the nitrogen-rich of PEI, the relative ECL quantum efficiency of the proposed N-Ti3C2 QDs was calculated to be 1.5 times higher than our previous reported N-Ti3C2 QDs that utilizing ethylenediamine as nitrogen source. Therefore, taking the advantage of the improved moving freedom and area of the interparticle DNA walker and high ECL efficiency of the proposed N-Ti3C2 QDs, the fabricated ECL biosensor for MUC1 determination displayed a low detection limit with a wide linear range, which provides a potential feasible method for other protein sensitive assay.

An In Vitro Comparison of Costimulatory Domains in Chimeric Antigen Receptor T Cell for Breast Cancer Treatment.

Adoptive cellular therapy with chimeric antigen receptor (CAR) T cells has emerged as a potential novel treatment for various cancers. In this study, we have generated CAR T cells targeting mucin-1 (MUC1), which is an aberrantly glycosylated antigen overexpressed on breast cancer cells. Two different signaling domains, including CD28 and 41BB, were incorporated and directly compared the superiority of different costimulatory signals. Two different CAR MUC1 constructs were transduced into primary T cells and evaluated their characteristics and antitumor activities against MUC1+ cancer cells. CAR MUC1 T cells showed high transduction efficiency and antigen specificity toward MUC1+ cancer cell lines and primary breast cancer cells. When coculturing with target cells, the transduced cells exhibited potent antitumor activity in vitro and secrete proinflammatory cytokines. Upon antigen stimulation, incorporation of the 41BB signaling domain was able to improve T cell proliferation and reduce surface PD1 expression and the upregulation of suppressive cytokines, when compared with CAR MUC1 containing the CD28 domain. Our findings show that CAR T cell targeting MUC1 can be effective against MUC1+ breast cancer cell and support the further development of CAR MUC1 T cells containing 41BB signaling in preclinical and clinical studies of breast cancer treatment.

Programmable bidirectional dynamic DNA nano-device for accurate and ultrasensitive fluorescent detection of trace MUC1 biomarker in serums

Accurate and ultrasensitive detection of biomarkers is significance for the diagnosis of diseases at early stage. For this purpose, we herein develop a bidirectional dynamic DNA nano-device for amplified fluorescent detection of tumor marker of mucin 1 (MUC1). The nano-device is constructed by immobilizing two sets of DNA cascade catalytic probes on two opposite directions of a single-stranded DNA tracker to limit probe reactants to a compact space. Once target MUC1 binds to the aptamer sequence, the initiator DNA locked in the duplex DNA substrate can be released to induce DNA-initiated cascade hybridization reactions (DCHRs) simultaneously in two opposite directions along the tracker DNA, accompanying the displacement of two quencher labeled-DNA intermediate initiators to facilitate successively execution of DCHRs on the DNA nano-devices, which results in the separation of fluorophore (FAM) and quencher (Dabycl) to produce substantially recovered fluorescent signals for rapid and sensitive detection of MUC1 with a detection limit down to 0.18 pM. In addition, this strategy also exhibits high selectivity against other interfering proteins and potential application capacity in real serum samples, indicating its promising application prospects in disease diagnosis and treatment.

Abstract 5423: Translational PK/PD/efficacy modeling and efficacious human dose prediction for a first-in-class MUC1-EGFR (M1231) bispecific antibody drug conjugate

Abstract Introduction: M1231 is a first-in-class bispecific antibody drug conjugate (ADC) targeting mucin-1 (MUC1) and epidermal growth factor receptor (EGFR), that is conjugated with a novel hemiasterlin-related microtubule inhibitor payload. Following dose, M1231 binds to EGFR and co-binds to tumor-associated hypoglycosylated MUC1, internalizes into the tumor cell and traffics to lysosomes where the payload is enzymatically released to affect cell viability. Methods: A Multi-scale Systems Pharmacology model was developed to account for the ADC’s disposition and interactions with the underlying physiologic system, resulting in intracellular payload release in tumor cells to drive tumor growth inhibition (TGI). An in vitro model quantified ADC internalization and lysosomal trafficking in the MUC1 and EGFR-expressing cancer cell lines MDA-MB-468 and OVCAR-3. TGI was assessed in mice bearing MUC1-tumors from the squamous NSCLC patient-derived xenograft model, LUX003. Pharmacokinetics in cynomolgus monkeys was modeled using a Target Mediated Drug Disposition model and was allometrically scaled to humans. Data Summary: All preclinical modeling results were integrated and scaled to simulate plasma M1231 concentrations to predict the human dose and corresponding exposures. Tumor stasis was estimated to begin at a dose of 2.4 mg/kg every 3 weeks (Q3W) with a maximum tumor regression achieved at a dose of 4.3 mg/kg Q3W. Conclusions: The quantitative systems pharmacology model-based efficacious dose prediction range of 2.4 mg/kg to 4.3 mg/kg dosed Q3W informed the design of the ongoing M1231 first-in-human trial (NCT04695847). Citation Format: Anup Zutshi, Berend Neuteboom, Seema Kumar, Willem Sloot, Christine Knuehl, Julia Dotterweich, Jianguo Ma, Christiane Amendt, Karthik Venkatakrishnan, Taeshin Park, John Pappas, Kyoung-Ae Kim. Translational PK/PD/efficacy modeling and efficacious human dose prediction for a first-in-class MUC1-EGFR (M1231) bispecific antibody drug conjugate [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5423.

Methylation-Mediated Silencing of MicroRNA-497 Promotes Breast Cancer Progression Through Up-Regulation of Mucin1.

BackgroundPotential anti-tumor effects of microRNA-497 (miR-497) have been highlighted in various malignancies including breast cancer. However, little is known about the function of miR-497 and its putative target mucin1 (MUC1) in breast cancer. The present study explored how miR-497 regulates breast cancer progression in a MUC1-dependent manner.MethodsExpression of miR-497 and MUC1 was determined in breast cancer tissues and cells. Methylation specific polymerase chain reaction was used to measure the methylation status of CpG islands of miR-497 promoter, while chromatin immunoprecipitation assay was used to detect recruitment of methyltransferase to the promoter region of miR-497. Alteration in expression of miR-497 (overexpression) and MUC1 (up- and down-regulation) was performed to examine their roles in breast cancer biology in vitro and in vivo. The binding affinity between miR-497 and MUC1 was investigated through a bioinformatics database and dual luciferase reporter gene assay.ResultsMiR-497 was down-regulated and MUC1 was up-regulated in breast cancer tissues and cell lines. Besides, methylation induced a down-regulation of miR-497 in breast cancer. The bioinformatics analysis and dual luciferase reporter gene assay indicated that miR-497 targeted MUC1. Overexpression of miR-497 inhibited breast cancer cell proliferation and invasion and promoted the apoptosis of breast cancer cells by down-regulating MUC1. The inhibitory action of miR-497 on tumor growth was validated in vivo.ConclusionIn conclusion, miR-497 down-regulated MUC1 expression and subsequently suppressed breast cancer progression, highlighting miR-497 to be a potential biomarker and therapeutic target for breast cancer therapy.

Silver nanoclusters-based fluorescent biosensing strategy for determination of mucin 1: Combination of exonuclease I-assisted target recycling and graphene oxide-assisted hybridization chain reaction

A novel label-free fluorescent biosensing strategy was described for the sensitive detection of mucin 1 (MUC1). It consisted of an M-shaped aptamer probe for exonuclease I (Exo I)-assisted target recycling (EATR) amplification, and two AgNCs-hairpin probes for graphene oxide (GO)-assisted hybridization chain reaction (HCR) amplification. Based on the specificity of aptamer-target recognition, the addition of MUC1 caused a conformational change in the M-shaped aptamer probe, which was split into a MUC1-P3 complex and a P1-P2 duplex. Exo I then catalyzed the cleavage of aptamer sequence P3 from the MUC1-P3 complex and released the target MUC1. The released target MUC1 was free to bind with a new M-shaped probe to perform EATR amplification. Furthermore, the P1-P2 duplex with three single-stranded arms can act as a primer to initiate HCR between hairpin probes AgNCs-H1 and AgNCs-H2. In the process of HCR, two AgNCs-hairpins were autonomously cross-opened, generating long linear double-stranded nanowires containing large numbers of AgNCs. These nanowires cannot be quenched by GO due to the weak affinity between the long double-stranded DNA and GO, thereby retaining a strong fluorescent signal indicative of the concentration of MUC1. With these designs, in addition to an extremely low detection limit of 0.36 fg mL−1, the method exhibited an acceptable linear response to detect MUC1 from 1 fg mL−1 to 1 ng mL−1. Additionally, this method could be exerted with a high degree of success to detect MUC1 in diluted human serum with satisfactory results.

Development of a fluorescence immunoassay based on X-Ti-Zn nanocomposite for the detection of MUC1 biomarker

Synthesis of novel nanomaterials with fluorescence or quenching properties is a key factor for development of fluorescent-based assays as one the most widely used detection methods. In the present study, Ag-Ti-Zn nanocomposite as a new class of zinc-based nanocomposites with fluorescence quenching property was developed for the detection of Mucin 1 (MUC1). Prepared nanocomposite was characterized by atomic force microscopy (AFM), Raman spectroscopy, UV–vis spectroscopy and photoluminescence spectroscopy. In the absence of target MUC1, Ag-Ti-Zn nanocomposite was able to quench fluorescence of Rhodamine-B (RhB) dye while they were in close proximity of each other. The addition of MUC1 antibody to the RhB/Ag-Ti-Zn nanocomposite system further decreased fluorescence intensity of RhB. Specific binding of the MUC1 to antibody induced precipitation of nanocomposite and recovered fluorescence of RhB. Under optimized conditions, the limit of detection (LOD) for MUC1 was as low as 0.02 U/mL with linear detection rage from 0.05 to 3 U/ mL. The method was very sensitive, selective and simple for the detection of MUC1. The results exhibited that Ag-Ti-Zn nanocomposite can be used as a common quencher and sensing platform to design highly sensitive fluorescent-based assays.

MUC1 confers radioresistance in head and neck squamous cell carcinoma (HNSCC) cells

ABSTRACT Mucin 1 (MUC1), a transmembrane glycoprotein, has shown to be as the possible prognostic marker to predict the risk of aggressive head and neck squamous cell carcinoma (HNSCC). In the present study, we investigated the effect of MUC1 in HNSCC cells and the response to X-ray irradiation (IR). Here, we examined the impact of MUC1 overexpression or downexpression on clonogenic survival and apoptosis in response to X-ray irradiation (IR). Radioresistance and radiosensitivity were also observed in HNSCC cells that are MUC1 overexpression and MUC1 downexpression. This enhanced resistance to IR in MUC1-overexpressing cells is primarily due to increased the number of radiation-induced γH2AX/53BP1-positive foci and DNA double-strand break (DSB) repair kinetics. MUC1 overexpression repaired more than 90% of DSBs after 2 Gy radiation by 24 h compared to the empty vector overexpressing cells with less than 50% of DSB repair. However, MUC1 downexpression repaired less than 20% of DSBs compared to the empty vector-overexpresing cells. MUC1 overexpression inhibited proapoptotic protein expression, such as caspase-3, caspase-8, and caspase-9, and induced antiapoptotic protein Bcl-2, followed by resistance to IR-induced apoptosis. Our results showed that targeting MUC1 may be as a promising strategy to counteract radiation resistance of HNSCC cells.

Sandwich immunoassay coupled with isothermal exponential amplification reaction: An ultrasensitive approach for determination of tumor marker MUC1

An ultrasensitive strategy based on sandwich immunoassay coupled with isothermal exponential amplification reaction (IMEXPAR) is proposed for the determination of tumor protein Mucin 1 (MUC1). An immuno-PCR plate was prepared from modification of the primary MUC1-antibody (Ab1) onto the inner-well of the PCR plate. A biotinylated secondary MUC1-antibody tagged with the biotinylated EXPAR primer (P-Ab2) was prepared through biotin-streptavidin reaction. In the presence of target MUC1, sandwich-type combinations were specifically formed in the immuno-PCR plate. With further addition of amplification template, polymerase and nicking enzyme, EXPAR was specifically triggered, producing numerous primer replica in minutes, and greatly enhanced fluorescence of SYBR Green I. The proposed strategy has a good linear relationship with the logarithm of the MUC1 concentration ranging from 3 pM to 3 nM with a limit of detection of 1.63 pM (S/N = 3), which is two orders of magnitude lower than those of other methods. Owing to the specificity of immuno-reaction and EXPAR, the selectivity of the strategy is favorable, even if for the homologous protein. The proposed strategy was further applied for the MUC1 determination in human serum, and a satisfactory recovery range of 98.7%–105.3% was obtained. The strategy can be facilely extended to the ultrasensitive determination of various proteins.

Ruthenium complex doped metal-organic nanoplate with high electrochemiluminescent intensity and stability for ultrasensitive assay of mucin 1

In this study, a novel Ru(bpy)2(dcbpy)-doped two-dimensional (2D) metal-organic framework (MOF) nanoplate (Ru@Zr12-BPDC, BPDC = 4,4′-biphenyldicarboxylate, bpy = 2,2'-bipyridine, H2dcbpy = 2,2'-bipyridine-5,5'-dicarboxylic acid) with outstanding ECL performance was synthesized via a mixed ligand strategy. The matching lengths between Ru(bpy)2(dcbpy) and BPDC ligands allowed the Ru(bpy)2(dcbpy) to bridge two Zr12 clusters of Ru@Zr12-BPDC nanoplate through strong coordination bonds, which not only resulted in the increase of the immobilization amount of Ru(bpy)2(dcbpy), but also prevented the leakage of the Ru(bpy)2(dcbpy) effectively. Furthermore, the high porosity and ultrathin nature of 2D Ru@Zr12-BPDC nanoplate not only allowed the electrochemical activation of the internal and external Ru(bpy)2(dcbpy), but also greatly shortened the diffusion paths of the ions, electrons, coreactant (tripropylamine, TPrA) and coreactant intermediates (TPrA• and TPrA•+), which made more interior Ru(bpy)2(dcbpy) could be excited and thus led to the increasing utilization ratio of the luminophores. Because of the above merits, the Ru@Zr12-BPDC nanoplate exhibited high and stable ECL emission, and thus was employed to construct an aptasensor for ultrasensitive detection of mucin 1 (MUC1). To further enhance the sensitivity of the proposed aptasensor and amplify signal, the target MUC1 was converted into the H1-H2 duplex with the assistance of H1, H2 and MUC1-catalyzed hairpin assembly (CHA). As expected, the proposed aptasensor exhibited remarkable stability, excellent sensitivity, high sensitivity and displayed a wide liner range from 1 fg/mL to 10 ng/mL with a low detection limit of 0.14 fg/mL. To our knowledge, it is the first example of the construction of an ECL biosensor based on ruthenium complex doped 2D MOF nanoplates. This study demonstrated that the use of ruthenium complex doped MOF nanoplates to increase the ECL stability and intensity is an efficient strategy for the construction of highly stable and sensitive ECL biosensors, and therefore may extend the applications of 2D MOF nanoplates in ECL and bioanalysis fields.

Enhanced photoelectrochemical sensing for MUC1 detection based on TiO2/CdS:Eu/CdS cosensitized structure

A novel ultrasensitive photoelectrochemical aptasensing was developed to detect mucin 1 (MUC1) based on TiO2/CdS:Eu nanocomposite. The TiO2/CdS:Eu nanocomposite modified fluorine-doped tin oxide (FTO) electrode was prepared by successive ionic layer adsorption and reaction technique (SILAR) and then was employed as matrix for immobilization of SH-DNA (DNA1). After 6-mercaptohexanol (MCH) blocked unbound sites of the electrode surface, the MUC1 aptamer(DNA2) was introduced, allowing MUC1 aptamer to hybridize with DNA1. Subsequently, CdS-COOH QDs as signal amplification elements were successively labeled on the terminal of MUC1 aptamer. In this case, the target MUC1 detection was based upon the photocurrent change originated from quantities change of the signal amplification elements CdS. In the presence of MUC1, the MUC1 aptamer was specifically bound with MUC1, and meanwhile MUC1/DNA2/CdS was released from the electrode surface, resulting in significantly decreased photocurrent intensity. The novel ultrasensitive photoelectrochemical aptasensing presented a highly selective, sensitive and reproductive detection of MUC1 from 1 nM to 5 μM with a lower detection limit of 0.3 nM. At the same time, it also provided a good specificity, reproducibility, stability and promising strategy to detect a variety of other protein targets for early clinical diagnosis of different diseases.

Enzyme-free Target Recycling and Double-Output Amplification System for Electrochemiluminescent Assay of Mucin 1 with MoS2 Nanoflowers as Co-reaction Accelerator.

In this work, a sensitive electrochemiluminescent assay of mucin 1 (MUC1) was developed with the advantages of target recycling amplification strategy and effective MoS2 nanoflower (MoS2 NF)-based signal probe. Briefly, the target MUC1 triggered enzyme-free recycling and a double-output amplification process was executed to acquire masses of single-stranded DNA as a mimic target, which further participated in the catalytic hairpin assembly process for signal amplification. Meanwhile, MoS2 NFs were prepared as an effective co-reaction accelerator, which not only possessed excellent catalytic performance for H2O2 decomposition to largely enhance the luminous intensity of N-(aminobutyl)- N-(ethylisoluminol) (ABEI)-H2O2 electrochemiluminescence system but also offered a desirable platform for ABEI-functionalized Ag nanoparticles (ABEI-Ag complexes) loading via Ag-S binding. The experimental results showed the proposed aptasensor had a good linear relationship in the range of 1 fg/mL to 10 ng/mL for MUC1 detection and the limit of detection was 0.58 fg/mL (S/N = 3). In addition, the aptasensor had nice stability and selectivity and huge potential to be applied in clinical research.

Abstract LB-092: Induction of IL-17-mediated cellular immunity targeting MUC1 through vaccination

Abstract Prophylactic vaccination targeting tumor-associated antigens (TAA) holds the potential to prevent the development of cancer. Clinical evidence has demonstrated a protective effect of cellular immunity against the mucin 1 (MUC1) TAA, expressed by ∼90% of mammary tumors. Nevertheless, conventional vaccination strategies have failed to prevent breast cancer due to the induction of terminally differentiated, short-lived Th1 cells of limited functionality. By contrast, IL-17-producing T cells, including CD4+ Th17 and CD8+ Tc17 subtypes, account for pathogenic agents that escape Th1-mediated control and are thus endowed with specialized, non-redundant mechanisms to promote inflammation at the cell and tissue levels. Recent studies have confirmed that signaling through the inducible T-cell co-stimulator (ICOS) pathway is sufficient to polarize T cells towards IL-17-expression in vitro, providing a novel mechanism to induce Th17/Tc17-mediated immunity against tumors in vivo. To generate IL-17-mediated antitumor immunity through vaccination, we examined the immunogenicity of combinations of adenoviral vectors encoding MUC1, ICOS ligand (ICOS-L) and the cytokines IL-6 and IL-23, implicated in Th17 development and expansion, respectively. Vaccination against MUC1, in the presence or absence of IL-17-polarizing conditions, overcame immune tolerance in transgenic mice recognizing MUC1 as a “self” antigen, inducing robust MUC1-specific intracellular IFNγ production from CD8+ T cells detected by FACS. Co-administration of vectors encoding MUC1 and ICOS-L, with or without vector-derived IL-6 and/or IL-23, induced significant IL-17 production in response to mitogen from both CD4+ and CD8+ T cells compared to MUC1 vaccination alone. Interestingly, ICOS-L augmentation resulted in the upregulation of the master Th17 transcription factor RORγt in CD4+, but not CD8+ T cells. Additionally, administration of ICOS-L, with or without IL-6 and/or IL-23, increased levels of granzyme B and CD107a in both CD4+ and CD8+ T cell populations following MUC1 vaccination, suggesting IL-17-polarization enhances T cell cytotoxicity. Studies are currently ongoing to evaluate the efficacy of IL-17-polarizing or conventional MUC1-targeting vaccination to protect MUC1-tolerant, transgenic mice against an orthotopic challenge of MUC1-expressing EO771 cells and findings will be presented at the AACR Annual Meeting. To the best of our knowledge, this study represents the first demonstration that IL-17-polarized immunity may be induced through vaccination, providing a novel platform for the generation of MUC1-specific immunity for the prevention of breast cancer. Citation Format: Amanda C. LaRue, Rebecca Carrell, Lindsay T. McDonald, Dayvia A. Russell, Stephen P. Ethier, Adam C. Soloff. Induction of IL-17-mediated cellular immunity targeting MUC1 through vaccination. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-092.

Electrochemiluminescence Aptasensor Based on Cascading Amplification of Nicking Endonuclease-Assisted Target Recycling and Rolling Circle Amplifications for Mucin 1 Detection

In this work, a novel and sensitive electrochemiluminescence (ECL) aptasensor for mucin 1 (MUC1) was designed based on cascading amplification of nicking endonuclease-assisted target recycling and rolling circle amplifications (RCA). In the presence of target MUC1, Nt.BbvC I-assisted target recycling process generated massive single-stranded primers which were available for subsequent RCA reactions on the as-prepared sensing interface. With rationally sequence designed, massive long DNA molecules with multiple G-quadruplex units were formed to generate the G-quarduplex/hemin complexes in the presence of hemin, which could enhance the ECL efficiency of luminol-H2O2 system due to its strongly catalytic performance toward the coreactant H2O2. The proposed aptasenor for MUC1 detection showed a limit of detection down to 0.71fg/mL and a wide linear range from 10fg/mL to 10ng/mL with acceptable stability, sensitivity, and possessed promising potential applications in clinical diagnostics. More significantly, it would be available for generalizing the proposed strategy to detect a series of other relative biomarkers.

Upregulation of oncogenic MUC1 by the EMT transcription factor brachyury mediates immunotherapy-resistance in tumor cells

Epithelial-mesenchymal transition (EMT) is a molecular and cellular program in which epithelial cells lose their well-differentiated phenotype and adopt mesenchymal characteristics. This process occurs during the progression of cancer to metastatic disease, and is also associated with recurrence as conventional therapies fail to eliminate the minority of resistant tumor cells that have undergone EMT. Brachyury is a transcription factor that controls EMT during development, and we have shown that brachyury is aberrantly overexpressed in various human cancers where it promotes tumor cell EMT, metastatic dissemination, and resistance to conventional therapies. Further, we have recently reported that very high expression of brachyury can protect against caspase-dependent immune cell-mediated cytotoxicity. In seeking to elucidate mechanisms of immunotherapy resistance, we have discovered a novel association between brachyury and mucin-1 (MUC1). MUC1 is overexpressed in the majority of carcinomas, and it has been shown to mediate oncogenic signaling, induce EMT, increase stem cell characteristics, and confer resistance to genotoxic agents. Here, we report a novel positive association between brachyury and MUC1 in both cancer cell lines and patient tumor tissues, in which brachyury was found to upregulate MUC1 expression at the level of transcript stability. Targeting MUC1 by siRNA-based gene silencing was found to abrogate the resistance of brachyury-expressing tumor cells to killing by recombinant tumor necrosis-related apoptosis-inducing ligand (TRAIL) and to lysis by natural killer (NK) cells. These studies confirm a protective role for MUC1 as a mediator of immunotherapy-resistance in brachyury-expressing tumor cells, and demonstrate that targeting MUC1 restores the susceptibility of mesenchymal tumor cells to immune attack.

Antitumor effect of adenoviral vector prime protein boost immunity targeting the MUC1 VNTRs

Mucin 1 (MUC1) is a tumor-associated antigen that is overexpressed in several adenocarcinomas. However, clinical trials with MUC1 showed that MUC1 is a relatively poor immunogen in humans. In view of the low immunogenicity of this protein vaccine, we designed a method based on an immunoadjuvant and immunization strategy to enhance the cellular immune response to this protein vaccine. DDA/MPL has been evaluated as an adjuvant to induce strong immunity for the tuberculosis vaccine. However, its adjuvant role combined with the vaccine targeting MUC1 in malignant carcinomas has not previously been reported. Our previous study showed that adenovirus prime protein boost vaccination could significantly enhance the cellular immunity and antitumor efficacy. In our study, we used MUC1 VNTRs as the target of cancer vaccine and DDA/MPL as the adjuvant to enhancing the cellular immunity of recombinant MUC1 protein vaccine, and an AD-9M adenoviral vector prime-recombinant protein and DDA/MPL boost (designated MUC-1 VPP vaccine) strategy was studied to enhance the antitumor efficacy. The results demonstrated that antigen-specific IFN-γ-secreting T cells were increased by 2-fold, and cytotoxic T lymphocytes (CTLs) were induced effectively when the protein vaccine was combined with the DDA/MPL adjuvant. Moreover, the vaccination induced nearly 60% inhibition of the growth of B16 melanoma in mice and prolonged the survival of tumor-bearing mice. The inhibition was correlated with the specific immune responses induced by the MUC1 VPP vaccine. The data suggested that DDA/MPL-adjuvant MUC-1 VPP vaccine may be developed into effective tumor vaccines for melanomas and possibly for other tumors expressing MUC1 protein.

A study of autologous dendritic cell therapy targeting mucin-1 for treatment of patients with epithelial ovarian cancer in first remission.

TPS5113 Background: Cvac (Cvac) is an autologous dendritic cell immune stimulant targeting mucin-1 positive tumors. A phase IIa study demonstrated potential for treating mucin-1 positive epithelial ovarian carcinoma (EOC). A phase IIb study was designed to determine on a pilot basis if Cvac could demonstrate an effect on progression free and overall survival in patients with EOC in first or second remission. The current study, CANVAS (CANcer VAccine Study) is evaluating Cvac treatment of EOC patients in first remission after surgery and standard chemotherapy, with at least three cycles of platinum and taxane therapy. Endpoints include progression free survival, overall survival, safety evaluation and quality of life scores. Methods: Patients are eligible if they have stage III or IV mucin-1 positive EOC and have had optimal cytoreductive (<1cm residual disease) surgery. Apheresis to obtain immature dendritic cells occurs prior to chemotherapy. Cvac is manufactured during chemotherapy. Patients remain eligible for the study if they have a complete response to standard chemotherapy after surgery. Cvac or placebo is dosed every four weeks for three doses, and every twelve weeks for three additional doses. 1000 patients will be randomized to Cvac or placebo in 22 countries, with 800 expected to receive vaccine or placebo. CT or MRI imaging determines progression, and is centrally read prior to inclusion and at each timepoint. Recruitment commenced January 2012. Over 100 centers are participating in the study globally, with majority in Europe. Enrollment is expected to take 18-24 months. An interim analysis is planned after 400 PFS events, and final analysis after 620 PFS events. Observation of 620 events would have approximately 90% power to detect a hazard ratio of 0.77 in PFS at a significance level of p=0.049 (approximately a 30% increase in median PFS in the Cvac group compared with the placebo group).