Fig 2 - uploaded by Jerry Nickles
Content may be subject to copyright.
Flow cytometry analysis of ALT-836-Fab at different concentrations (1 μg/mL and 5 μg/ml) in MDA-MB-231 (TF positive) and MDA-MB-435 (TF negative) cells. All data are repeated 3 times per group.
Source publication
To date, there is no effective therapy for triple-negative breast cancer (TNBC), which has a dismal clinical outcome. Upregulation of tissue factor (TF) expression leads to increased patient morbidity and mortality in many solid tumor types, including TNBC. Our goal was to employ the Fab fragment of ALT-836, a chimeric anti-human TF mAb, for PET im...
Contexts in source publication
Context 1
... in vivo studies, flow cytometry was performed to examine the targeting efficiency and specificity of ALT-836-Fab in vitro. After treatment with FITC-ALT-836-Fab at a low concentration (1 μg/mg), the fluorescence signal from MDA-MB-231 cells (TF positive) exhibited a 25-fold increase over the negative group, whereas that from MDA-MB-435 cells (TF negative) only exhibited 4-fold increase (n = 3; Fig. 2). This result clearly demonstrated the TF specificity of ...
Context 2
... in vivo studies, flow cytometry was performed to examine the targeting efficiency and specificity of ALT-836-Fab in vitro. After treatment with FITC-ALT-836-Fab at a low concentration (1 μg/mg), the fluorescence signal from MDA-MB-231 cells (TF positive) exhibited a 25-fold increase over the negative group, whereas that from MDA-MB-435 cells (TF negative) only exhibited 4-fold increase (n = 3; Fig. 2). This result clearly demonstrated the TF specificity of ...
Context 3
... in vivo studies, flow cytometry was performed to examine the targeting efficiency and specificity of ALT-836-Fab in vitro. After treatment with FITC-ALT-836-Fab at a low concentration (1 μg/mg), the fluorescence signal from MDA-MB-231 cells (TF positive) exhibited a 25-fold increase over the negative group, whereas that from MDA-MB-435 cells (TF negative) only exhibited 4-fold increase (n = 3; Fig. 2). This result clearly demonstrated the TF specificity of ...
Similar publications
The present study investigated the feasibility of improving the tumor‑targeting efficacy and decreasing the toxicity of liposomal cabazitaxel (Cab) with aptamer modification. The process involved preparing aptamer (TLS1c)‑modified liposomes and studying the behavior of the liposomes in vitro and in vivo. TLS1c as an aptamer, which has high specific...
Manganese has been used in tumor imaging for their ability to provide T1-weighted MRI signal. Recent research find Mn²⁺ can induce activation of the stimulator of interferon gene (STING) pathway to create an active and favorable tumor immune microenvironment. However, the direct injection of Mn²⁺ often results in toxicity. In this study, we develop...
Simple Summary
Chemotherapy has been widely used in clinic to treat various types of tumors, although severe side effects have been the most critical limitation of this treatment modality. For a long time, it has been believed that toxicity derives from the apoptosis of normal cells induced by the use of chemotherapeutic drugs due to their off-targ...
We demonstrate that Blautia coccoides JCM1395T has the potential to be used for tumor-targeted live bacterial therapeutics. Prior to studying its in vivo biodistribution, a sample preparation method for reliable quantitative analysis of bacteria in biological tissues was required. Gram-positive bacteria have a thick outer layer of peptidoglycans, w...
Introduction:
Cytokine-based products are gaining importance for cancer immunotherapy. L19-TNF is a clinical-stage antibody-cytokine fusion protein that selectively accumulates to tumors and displays potent anticancer activity in preclinical models. Here, we describe an innovative approach to transiently inhibit off-target toxicity of L19-TNF, whi...
Citations
... Antibody fragments, on the other hand, are smaller (50 kDa for antigen-binding fragment, Fab), with shorter biological half-life (faster clearance rates), reduced immunogenicity [16], decreased potential for accumulation in the extravascular space, and maintenance of target specificity of antibodies. The main caveat however is the decreased binding affinity compared to full antibodies [17]. ...
Purpose
Invasive fungal diseases, such as pulmonary aspergillosis, are common life-threatening infections in immunocompromised patients and effective treatment is often hampered by delays in timely and specific diagnosis. Fungal-specific molecular imaging ligands can provide non-invasive readouts of deep-seated fungal pathologies. In this study, the utility of antibodies and antibody fragments (Fab) targeting β-glucans in the fungal cell wall to detect Aspergillus infections was evaluated both in vitro and in preclinical mouse models.
Methods
The binding characteristics of two commercially available β-glucan antibody clones and their respective antigen-binding Fabs were tested using biolayer interferometry (BLI) assays and immunofluorescence staining. In vivo binding of the Zirconium-89 labeled antibodies/Fabs to fungal pathogens was then evaluated using PET/CT imaging in mouse models of fungal infection, bacterial infection and sterile inflammation.
Results
One of the evaluated antibodies (HA-βG-Ab) and its Fab (HA-βG-Fab) bound to β-glucans with high affinity (KD = 0.056 & 21.5 nM respectively). Binding to the fungal cell wall was validated by immunofluorescence staining and in vitro binding assays. ImmunoPET imaging with intact antibodies however showed slow clearance and high background signal as well as nonspecific accumulation in sites of infection/inflammation. Conversely, specific binding of [⁸⁹Zr]Zr-DFO-HA-βG-Fab to sites of fungal infection was observed when compared to the isotype control Fab and was significantly higher in fungal infection than in bacterial infection or sterile inflammation.
Conclusions
[⁸⁹Zr]Zr-DFO-HA-βG-Fab can be used to detect fungal infections in vivo. Targeting distinct components of the fungal cell wall is a viable approach to developing fungal-specific PET tracers.
... FVII is an enzyme primarily synthesized and secreted by hepatocytes that is involved in the extrinsic coagulation cascade [150,151]. The active form of this complex (TF-FVII) is upregulated in many solid tumors, leading to thrombin generation and hemostasis and resulting in cancer cell signaling, inhibition of apoptosis, promotion of cell migration, tumor angiogenesis, and metastatic spread through thrombin generation and PAR1 signaling [150][151][152][153]. TF is highly expressed in several human TNBC cell lines, such as MDA-MB-231, MDA-MB-468, and HCC-1806 [154]. High levels of TF have been found to contribute to disease progression in TNBC patients [149][150][151]155], and TF levels can act as a prognostic marker, as high TF expression usually contributes to a decreased overall survival rate in TNBC patients [153]. ...
... Factor VII, the natural ligand to TF, can be radiolabeled to allow for specific imaging of TF, and as such could be used in imaging and therapy radionuclides for TNBC [153,154]. As for other tumor markers, assessing tumor TF status based on tissue sampling may be limited by both the invasive nature of biopsy and by tumor heterogeneity, especially in the context of metastatic disease [153,154]. ...
... Factor VII, the natural ligand to TF, can be radiolabeled to allow for specific imaging of TF, and as such could be used in imaging and therapy radionuclides for TNBC [153,154]. As for other tumor markers, assessing tumor TF status based on tissue sampling may be limited by both the invasive nature of biopsy and by tumor heterogeneity, especially in the context of metastatic disease [153,154]. Several pre-clinical studies have explored the utility of radi-olabeling TF to evaluate TNBC. The first TF PET imaging was performed on subcutaneous xenograft mouse models using a 64 Cu-labelled anti-TF antibody, 64 Cu-NOTA-ALT-836. ...
One out of eight women will be affected by breast cancer during her lifetime. Imaging plays a key role in breast cancer detection and management, providing physicians with information about tumor location, heterogeneity, and dissemination. In this review, we describe the latest advances in PET/CT imaging of breast cancer, including novel applications of 18F-FDG PET/CT and the development and testing of new agents for primary and metastatic breast tumor imaging and therapy. Ultimately, these radiopharmaceuticals may guide personalized approaches to optimize treatment based on the patient’s specific tumor profile, and may become a new standard of care. In addition, they may enhance the assessment of treatment efficacy and lead to improved outcomes for patients with a breast cancer diagnosis.
... It indicates the excellent affinity and TF-specificity of this radiolabeled PET-tracer [219]. Several-fold greater tumor uptake of (64) Cu-NOTA-ALT-836-Fab compared to the blocking group and tumor models that failed to significantly express TF was seen in a serial PET imaging study of the MDA-MB-231 TNBC model [218]. Dual-targeted imaging agents have indicated enhanced targeting effectiveness rather than singletargeted entities. ...
Tissue factor (TF) is a protein that plays a critical role in blood clotting, but recent research has also shown its involvement in cancer development and progression. Herein, we provide an overview of the structure of TF and its involvement in signaling pathways that promote cancer cell proliferation and survival, such as the PI3K/AKT and MAPK pathways. TF overexpression is associated with increased tumor aggressiveness and poor prognosis in various cancers. The review also explores TF's role in promoting cancer cell metastasis, angiogenesis, and venous thromboembolism (VTE). Of note, various TF-targeted therapies, including monoclonal antibodies, small molecule inhibitors, and immunotherapies have been developed, and preclinical and clinical studies demonstrating the efficacy of these therapies in various cancer types are now being evaluated. The potential for re-targeting TF toward cancer cells using TF-conjugated nanoparticles, which have shown promising results in preclinical studies is another intriguing approach in the path of cancer treatment. Although there are still many challenges, TF could possibly be a potential molecule to be used for further cancer therapy as some TF-targeted therapies like Seagen and Genmab’s tisotumab vedotin have gained FDA approval for treatment of cervical cancer. Overall, based on the overviewed studies, this review article provides an in-depth overview of the crucial role that TF plays in cancer development and progression, and emphasizes the potential of TF-targeted and re-targeted therapies as potential approaches for the treatment of cancer.
... The antibodies are utilized in diagnosis and immunotherapy of cancer. To conceptualize the differential upregulation in expression level of tissue factor (TF) receptor and urokinase plasminogen activator receptor (uPAR) in TNBC, researchers validated the utility of anti-TF antibody labeled with Cu-64 (anti-TF-antibody-64Cu) using PET imaging in TNBC in vitro model [165]. LeBeau et al. detected NIR fluorophore and Indium-111 ( 111 In) labeled uPAR antibodies using optical and SPECT imaging, respectively [166]. ...
Breast cancer is the second most cancer across the globe with high incidence (11.6%) and mortality (6.6%) among women. Surgery, radiotherapy, chemotherapy, neoadjuvant, and adjuvant therapy are the best possible ways to manage the breast cancer. However, the undesirable effects of anticancer chemotherapy on healthy breast tissues and increasing drug resistance are the most important hurdles in defeating the breast cancer. Hence, the scope of targeting the tumor microenvironment, oncogenes, bio-nucleotides, and inhibition of metastasis has become emergent through nanomedicine and theranostic approaches. Also, the effective delivery and selective targeting of chemotherapeutic drugs to reach the desired site (receptor or target) of cancer are in emergent need and become possible through nanomedicines and theranostics. Therefore, this chapter focuses on the advancements in the delivery of formulated nanomedicines and theranostics for defeating breast cancer. The applications and challenges of nanomedicine delivery systems and theranostic approach for breast cancer treatment in clinical perspectives are also detailed here.
... Identifier: NCT01325558). A radiolabeled antibody fragment, 64 Cu-NOTA-ALT-836-Fab (Fab, Figure 2), was developed and shown to have an uptake level of about 4% ID/ g with T/B ratio of 2 at 24 h p. i. in MDA-MB-231 xenograft models (Shi et al., 2015). Therefore, targeting TF could be a potential way for imaging and therapy of TNBC. ...
Breast cancer is the most common cancer in women worldwide. The heterogeneity of breast cancer and drug resistance to therapies make the diagnosis and treatment difficult. Molecular imaging methods with positron emission tomography (PET) and single-photon emission tomography (SPECT) provide useful tools to diagnose, predict, and monitor the response of therapy, contributing to precision medicine for breast cancer patients. Recently, many efforts have been made to find new targets for breast cancer therapy to overcome resistance to standard of care treatments, giving rise to new therapeutic agents to offer more options for patients with breast cancer. The combination of diagnostic and therapeutic strategies forms the foundation of theranostics. Some of these theranostic agents exhibit high potential to be translated to clinic. In this review, we highlight the most recent advances in theranostics of the different molecular subtypes of breast cancer in preclinical studies.
... In another study, Robinson et al demonstrated that 1.665 MBq of 211 At-conjugated diabodies was close to the acute MTD in BALB/c nude mice based on body weight loss after administration. 45As various anti-TF mAbs have been successfully applied to ADC, 14-21 an antibody-photosensitizer conjugate,22 and a molecular imaging probe,24,27,28,46,47 TF is one of the attractive molecules for cancer targeting. In this study, we succeeded in preparing an anti-TF mAb labeled with 211 At. ...
Tissue factor (TF), the trigger protein of the extrinsic blood coagulation cascade, is abundantly expressed in various cancers including gastric cancer. Anti‐TF monoclonal antibodies (mAbs) capable of targeting cancers have been successfully applied to armed antibodies such as antibody‐drug conjugates (ADCs) and molecular imaging probes. We prepared an anti‐TF mAb, clone 1084, labeled with astatine‐211 (²¹¹At), as a promising alpha emitter for cancer treatment. Alpha particles are characterized by high linear energy transfer and a range of 50–100 µm in tissue. Therefore, selective and efficient tumor accumulation of alpha emitters results in potent antitumor activities against cancer cells with minor effects on normal cells adjacent to the tumor. Although the ²¹¹At‐conjugated clone 1084 (²¹¹At‐anti‐TF mAb) was disrupted by an ²¹¹At‐induced radiochemical reaction, we demonstrated that astatinated anti‐TF mAbs eluted in 0.6% or 1.2% sodium ascorbate (SA) solution were protected from antibody denaturation, which contributed to the maintenance of cellular binding activities and cytocidal effects of this mAb. Although body weight loss was observed in mice administered a 1.2% SA solution, the loss was transient and the radioprotectant seemed to be tolerable in vivo. In a high TF–expressing gastric cancer xenograft model, ²¹¹At‐anti‐TF mAb in 1.2% SA exerted a significantly greater antitumor effect than non‐protected ²¹¹At‐anti‐TF mAb. Moreover, the antitumor activities of the protected immunoconjugate in gastric cancer xenograft models were dependent on the level of TF in cancer cells. These findings suggest the clinical availability of the radioprotectant and applicability of clone 1084 to ²¹¹At‐radioimmunotherapy.
... In this approach, the antibody's primary role is to locate specific cell surface tumor markers or components located at the extracellular matrix, which is then recognized by the PET detection system [104]. Prominent evidence of this concept is the discovery of ATL-836 fragment antigen-binding (Fab) chimeric monoclonal antibody against human tissue factor (TF) [105]. The discovery of ATL-836 antibody provides a promising platform for future diagnostics and therapeutics of TNBC as TF, otherwise known as platelet tissue factor/factor III, engages a crucial role in the signaling of cancer cells (apoptosis inhibi-tion and cell migration promotion) and is found to be prominently presented on TNBC cells [106,107]. ...
Triple-negative breast cancer (TNBC) is an aggressive breast type of cancer with no expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2). It is a highly metastasized, heterogeneous disease that accounts for 10–15% of total breast cancer cases with a poor prognosis and high relapse rate within five years after treatment compared to non-TNBC cases. The diagnostic and subtyping of TNBC tumors are essential to determine the treatment alternatives and establish personalized, targeted medications for every TNBC individual. Currently, TNBC is diagnosed via a two-step procedure of imaging and immunohistochemistry (IHC), which are operator-dependent and potentially time-consuming. Therefore, there is a crucial need for the development of rapid and advanced technologies to enhance the diagnostic efficiency of TNBC. This review discusses the overview of breast cancer with emphasis on TNBC subtypes and the current diagnostic approaches of TNBC along with its challenges. Most importantly, we have presented several promising strategies that can be utilized as future TNBC diagnostic modalities and simultaneously enhance the efficacy of TNBC diagnostic.
... PET images revealed rapid uptake and retention of [ 64 Cu]Cu-NOTA-ALT-836-Fab in MDA-MB-231 xenografts with high tumor to background ratio. A limitation of this approach is the lower binding avidity of Fab fragments which is problematic if high absolute tumor uptake rather than high tumor contrast is required [29]. ...
Molecular imaging plays an increasingly important role in the diagnosis and treatment of different malignancies. Radiolabeled probes enable the visualization of the primary tumor as well as the metastases and have been also employed in targeted therapy and theranostic approaches. With breast cancer being the most common malignancy in women worldwide it is of special interest to develop novel targeted treatments. However, tumor microenvironment and escape mechanisms often limit their therapeutic potential. Addressing tumor stroma associated targets provides a promising option to inhibit tumor growth and angiogenesis and to disrupt tumor tissue architecture. This review describes recent developments on radiolabeled probes used in diagnosis and treatment of breast cancer especially in triple negative type with the focus on potential targets offered by the tumor microenvironment, like tumor associated macrophages, cancer associated fibroblasts, and endothelial cells.
... Tissue factor is a procoagulant protein that has emerged as a potential tumor marker since its overexpression is commonly observed in a variety of cancer types [14,31,32]. In this context, TF has been regarded as a target for tumor imaging and therapy [33][34][35]. The present study aimed to evaluate the ability of ixolaris, a potent tick-derived TF inhibitor [21], as a theranostic agent in experimental melanoma. ...
Tissue factor (TF), a blood coagulation protein, plays an important role in tumor growth, invasion, and metastasis. Ixolaris, a tick-derived non-immunogenic molecule that binds to TF, has demonstrated in vivo inhibitory effect on murine models of melanoma, including primary growth and metastasis. This work aimed to: I) develop an efficient and stable labeling technique of ixolaris with Iodine-131(131I); II) compare the biodistribution of 131I and 131I-ixolaris in tumor-free and melanoma-bearing mice; III) evaluate whether 131I-ixolaris could serve as an antimetastatic agent. Ixolaris radioiodination was performed using iodogen, followed by liquid paper chromatography. Labeling stability and anticoagulant activity were measured. Imaging studies were performed after intravenous administration of free 131I or 131I-ixolaris in a murine melanoma model employing the B16-F10 cell line. Animals were divided in three experimental groups: the first experimental group, D0, received a single-dose of 9.25 MBq of 131I-ixolaris at the same day the animals were inoculated with melanoma cells. In the second group, D15, a single-dose of 9.25 MBq of 131I-ixolaris or free 131I was applied into mice on the fifteenth day after the tumor induction. The third group, D1-D15, received two therapeutic doses of 9.25 MBq of 131I-ixolaris or 131I. In vitro studies demonstrated that 131I-ixolaris is stable for up to 24 h and retains its inhibitory activity on blood coagulation. Biodistribution analysis and metastasis assays showed that all treatment regimens with 131I-ixolaris were effective, being the double-treatment (D1/D15) the most effective one. Remarkably, treatment with free 131I showed no anti-metastatic effect. 131I-ixolaris is a promising theranostic agent for metastatic melanoma.
... tissue factor (TF) receptor and urokinase plasminogen activator receptor (uPAR) in TNBC, Shi et al.,[74] suggested and validated the application of the anti-TF antibody labeled with copper-64 (anti-TF-antibody-64 Cu) using PET imaging in in-vitro TNBC model. Le Beau et al.,[75] detected NIR fluorophore and Indium-111 ( 111 In) labelled uPAR antibodies using optical and SPECT imaging respectively. ...
