Showing papers by "Anja Feldmann published in 2023"
TL;DR: In this paper , the influence of palbociclib/fulvestrant on human CD3+ T cells and novel emerging T cell-based cancer immunotherapies was studied.
Abstract: The cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitor palbociclib is an emerging cancer therapeutic that just recently gained Food and Drug Administration approval for treatment of estrogen receptor (ER)-positive, human epidermal growth factor receptor (Her)2-negative breast cancer in combination with the ER degrader fulvestrant. However, CDK4/6 inhibitors are not cancer-specific and may affect also other proliferating cells. Given the importance of T cells in antitumor defense, we studied the influence of palbociclib/fulvestrant on human CD3+ T cells and novel emerging T cell-based cancer immunotherapies. Palbociclib considerably inhibited the proliferation of activated T cells by mediating G0/G1 cell cycle arrest. However, after stopping the drug supply this suppression was fully reversible. In light of combination approaches, we further investigated the effect of palbociclib/fulvestrant on T cell-based immunotherapies by using a CD3-PSCA bispecific antibody or universal chimeric antigen receptor (UniCAR) T cells. Thereby, we observed that palbociclib clearly impaired T cell expansion. This effect resulted in a lower total concentration of interferon-γ and tumor necrosis factor, while palbociclib did not inhibit the average cytokine release per cell. In addition, the cytotoxic potential of the redirected T cells was unaffected by palbociclib and fulvestrant. Overall, these novel findings may have implications for the design of treatment modalities combining CDK4/6 inhibition and T cell-based cancer immunotherapeutic strategies.
2 citations
TL;DR: In this article , an interdigitated gold nanowire-based impedance nanobiosensor was used to detect COVID-19-associated antigens and respective Abs appearing during and after infection.
Abstract: Detection of antigens and antibodies (Abs) is of great importance in determining the infection and immunity status of the population, as they are key parameters guiding the handling of pandemics. Current point-of-care (POC) devices are a convenient option for rapid screening; however, their sensitivity requires further improvement. We present an interdigitated gold nanowire-based impedance nanobiosensor to detect COVID-19-associated antigens (receptor-binding domain of S1 protein of the SARS-CoV-2 virus) and respective Abs appearing during and after infection. The electrochemical impedance spectroscopy technique was used to assess the changes in measured impedance resulting from the binding of respective analytes to the surface of the chip. After 20 min of incubation, the sensor devices demonstrate a high sensitivity of about 57 pS·sn per concentration decade and a limit of detection (LOD) of 0.99 pg/mL for anti-SARS-CoV-2 Abs and a sensitivity of around 21 pS·sn per concentration decade and an LOD of 0.14 pg/mL for the virus antigen detection. Finally, the analysis of clinical plasma samples demonstrates the applicability of the developed platform to assist clinicians and authorities in determining the infection or immunity status of the patients.
1 citations
TL;DR: In this paper , a first- or second-generation universal CAR (UniCAR) and target modules (TMs) were generated to target cells with so-called target modules which harbor an ErbB2 (HER2)-specific antibody domain for target cell binding and the E5B9 peptide recognized by the UniCAR.
Abstract: Chimeric antigen receptor (CAR)-engineered immune effector cells constitute a promising approach for adoptive cancer immunotherapy. Nevertheless, on-target/off-tumor toxicity and immune escape due to antigen loss represent considerable challenges. These may be overcome by adaptor CARs that are selectively triggered by bispecific molecules that crosslink the CAR with a tumor-associated surface antigen. Here, we generated NK cells carrying a first- or second-generation universal CAR (UniCAR) and redirected them to tumor cells with so-called target modules (TMs) which harbor an ErbB2 (HER2)-specific antibody domain for target cell binding and the E5B9 peptide recognized by the UniCAR. To investigate differential effects of the protein design on activity, we developed homodimeric TMs with one, two or three E5B9 peptides per monomer, and binding domains either directly linked or separated by an IgG4 Fc domain. The adaptor molecules were expressed as secreted proteins in Expi293F cells, purified from culture supernatants and their bispecific binding to UniCAR and ErbB2 was confirmed by flow cytometry. In cell killing experiments, all tested TMs redirected NK cell cytotoxicity selectively to ErbB2-positive tumor cells. Nevertheless, we found considerable differences in the extent of specific cell killing depending on TM design and CAR composition, with adaptor proteins carrying two or three E5B9 epitopes being more effective when combined with NK cells expressing the first-generation UniCAR, while the second-generation UniCAR was more active in the presence of TMs with one E5B9 sequence. These results may have important implications for the further development of optimized UniCAR and target module combinations for cancer immunotherapy.
1 citations
TL;DR: In this paper , the synthesis of nine small-molecule PD-L1 radiotracers with solubilizing sulfonic acids and a linker-chelator system was reported.
Abstract: Simple Summary PD-L1 plays a crucial role in the immune responses against cancer. Only around 30% of cancer patients respond to an anti-PD-L1 immune checkpoint inhibitor therapy. Noninvasive molecular imaging techniques such as positron emission tomography (PET) would allow identification of patients likely to respond. Here we report on the synthesis of nine PET radioligands targeting PD-L1, based on small-molecule inhibitors. We introduced a chelator for radiolabeling and water-soluble groups to aim for clearance through the kidneys. The compounds showed binding affinities toward PD-L1 in the lower nanomolar range and stability in vitro. Mouse experiments showed moderate accumulation in tumor tissue but mainly clearance through the liver. Additionally, the compounds showed unexpected long circulation times due to strong binding to albumin in blood. Nevertheless, our compounds are a starting point for further development of PD-L1 small molecule PET radiotracer to support therapy decisions. Abstract Noninvasive molecular imaging of the PD-1/PD-L1 immune checkpoint is of high clinical relevance for patient stratification and therapy monitoring in cancer patients. Here we report nine small-molecule PD-L1 radiotracers with solubilizing sulfonic acids and a linker–chelator system, designed by molecular docking experiments and synthesized according to a new, convergent synthetic strategy. Binding affinities were determined both in cellular saturation and real-time binding assay (LigandTracer), revealing dissociation constants in the single digit nanomolar range. Incubation in human serum and liver microsomes proved in vitro stability of these compounds. Small animal PET/CT imaging, in mice bearing PD-L1 overexpressing and PD-L1 negative tumors, showed moderate to low uptake. All compounds were cleared primarily through the hepatobiliary excretion route and showed a long circulation time. The latter was attributed to strong blood albumin binding effects, discovered during our binding experiments. Taken together, these compounds are a promising starting point for further development of a new class of PD-L1 targeting radiotracers.
TL;DR: In this article , the authors conjugated their previously described anti-PSCA monoclonal antibody (mAb) 7F5 with the bifunctional chelator CHX-A″-DTPA and subsequently radiolabeled it with the theranostic radionuclide 177Lu.
Abstract: Prostate specific membrane antigen (PSMA) is an excellent target for imaging and treatment of prostate carcinoma (PCa). Unfortunately, not all PCa cells express PSMA. Therefore, alternative theranostic targets are required. The membrane protein prostate stem cell antigen (PSCA) is highly overexpressed in most primary prostate carcinoma (PCa) cells and in metastatic and hormone refractory tumor cells. Moreover, PSCA expression positively correlates with tumor progression. Therefore, it represents a potential alternative theranostic target suitable for imaging and/or radioimmunotherapy. In order to support this working hypothesis, we conjugated our previously described anti-PSCA monoclonal antibody (mAb) 7F5 with the bifunctional chelator CHX-A″-DTPA and subsequently radiolabeled it with the theranostic radionuclide 177Lu. The resulting radiolabeled mAb ([177Lu]Lu-CHX-A″-DTPA-7F5) was characterized both in vitro and in vivo. It showed a high radiochemical purity (>95%) and stability. The labelling did not affect its binding capability. Biodistribution studies showed a high specific tumor uptake compared to most non-targeted tissues in mice bearing PSCA-positive tumors. Accordingly, SPECT/CT images revealed a high tumor-to-background ratios from 16 h to 7 days after administration of [177Lu]Lu-CHX-A″-DTPA-7F5. Consequently, [177Lu]Lu-CHX-A″-DTPA-7F5 represents a promising candidate for imaging and in the future also for radioimmunotherapy.
TL;DR: P6 as mentioned in this paper is a runtime verification framework for programmable switches that complements static analysis to automatically detect, localize, and patch software bugs in P4 programs, where bugs are reported via a violation of pre-specified expected behavior that is captured by P6.
Abstract: We introduce a runtime verification framework for programmable switches that complements static analysis. To evaluate our approach, we design and develop P6, a runtime verification system that automatically detects, localizes, and patches software bugs in P4 programs. Bugs are reported via a violation of pre-specified expected behavior that is captured by P6. P6 is based on machine learning-guided fuzzing that tests P4 switch non-intrusively, i.e., without modifying the P4 program for detecting runtime bugs. This enables an automated and realtime localization and patching of bugs. We used a P6 prototype to detect and patch existing bugs in various publicly available P4 application programs deployed on two different switch platforms, namely, behavioral model (bmv2) and Tofino. Our evaluation shows that P6 significantly outperforms bug detection baselines while generating fewer packets and patches bugs in large P4 programs, e.g., switch.p4 without triggering any regressions.
17 Jul 2023
TL;DR: In this article , the authors performed an in-depth study over six years and identified around 24k long-lived MOAS prefixes in IPv4 and 1.4k in IPv6 being announced in January 2023.
Abstract: BGP exchanges reachability information in the form of prefixes, which are usually originated by a single Autonomous System (AS). If multiple ASes originate the same prefix, this is referred to as a Multiple Origin ASes (MOAS) prefix. One reason for MOAS prefixes are BGP prefix hijacks, which are mostly short-lived and have been studied extensively in the past years. In contrast to short-lived MOAS, long-lived MOAS have remained largely understudied. In this paper, we focus on long-lived MOAS prefixes and perform an in-depth study over six years. We identify around 24k long-lived MOAS prefixes in IPv4 and 1.4k in IPv6 being announced in January 2023. By analyzing the RPKI status we find that more than 40% of MOAS prefixes have all origins registered correctly, with only a minority of MOAS having invalid origins. Moreover, we find that the most prominent CIDR size of MOAS prefixes is /24 for IPv4 and /48 for IPv6, suggesting their use for fine-grained traffic steering. We attribute a considerable number of MOAS prefixes to mergers and acquisitions of companies. Additionally, more than 90% of MOAS prefixes are originated by two origin ASes, with the majority of detected origin AS relations being customer-provider. Finally, we identify that the majority of MOAS users are IT companies, and just 0.9% of IPv4 MOAS and 6.3% of IPv6 MOAS prefixes are used for anycast.
TL;DR: In this article , a novel immunotherapeutic drug based on the SARS-CoV-2 entry receptor ACE2 was proposed for the treatment of the coronavirus disease 2019 (COVID-19) pandemic, which led to millions of infections and deaths worldwide.
Abstract: The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) led to millions of infections and deaths worldwide. As this virus evolves rapidly, there is a high need for treatment options that can win the race against new emerging variants of concern. Here, we describe a novel immunotherapeutic drug based on the SARS-CoV-2 entry receptor ACE2 and provide experimental evidence that it cannot only be used for (i) neutralization of SARS-CoV-2 in vitro and in SARS-CoV-2-infected animal models but also for (ii) clearance of virus-infected cells. For the latter purpose, we equipped the ACE2 decoy with an epitope tag. Thereby, we converted it to an adapter molecule, which we successfully applied in the modular platforms UniMAB and UniCAR for retargeting of either unmodified or universal chimeric antigen receptor-modified immune effector cells. Our results pave the way for a clinical application of this novel ACE2 decoy, which will clearly improve COVID-19 treatment.
24 Jul 2023
TL;DR: In this article , an out-of-band communication channel with Hypergiants that can be two or more AS hops away and, optionally, with the transit provider is proposed to better assign requests of end-users to appropriate Hypergiant servers.
Abstract: Today, there is an increasing number of peering agreements between Hypergiants and networks that benefit millions of end-user. However, the majority of Autonomous Systems do not currently enjoy the benefit of interconnecting directly with Hypergiants to optimally select the path for delivering Hypergiant traffic to their users. In this paper, we develop and evaluate an architecture that can help this long tail of networks. With our proposed architecture, a network establishes an out-of-band communication channel with Hypergiants that can be two or more AS hops away and, optionally, with the transit provider. This channel enables the exchange of network information to better assign requests of end-users to appropriate Hypergiant servers. Our analysis using operational data shows that our architecture can optimize, on average, 15% of Hypergiants' traffic and 11% of the overall traffic of networks that do not interconnect with Hypergiants. The gains are even higher during peak hours when available capacity can be scarce, up to 46% for some Hypergiants.
TL;DR: In this paper , a novel FMS-like tyrosine kinase 3 (FLT3)-directed UniCAR T-cell therapy, which is highly effective for in vitro killing of both AML cell lines and primary AML samples, was presented.
Abstract: Adaptor chimeric antigen receptor (CAR) T-cell therapy offers solutions for improved safety and antigen escape, which represent main obstacles for the clinical translation of CAR T-cell therapy in myeloid malignancies. The adaptor CAR T-cell platform 'UniCAR' is currently under early clinical investigation. Recently, the first proof of concept of a well-tolerated, rapidly switchable, CD123-directed UniCAR T-cell product treating patients with acute myeloid leukaemia (AML) was reported. Relapsed and refractory AML is prone to high plasticity under therapy pressure targeting one single tumour antigen. Thus, targeting of multiple tumour antigens seems to be required to achieve durable anti-tumour responses, underlining the need to further design alternative AML-specific target modules (TM) for the UniCAR platform. We here present the preclinical development of a novel FMS-like tyrosine kinase 3 (FLT3)-directed UniCAR T-cell therapy, which is highly effective for in vitro killing of both AML cell lines and primary AML samples. Furthermore, we show in vivo functionality in a murine xenograft model. PET analyses further demonstrate a short serum half-life of FLT3 TMs, which will enable a rapid on/off switch of UniCAR T cells. Overall, the presented preclinical data encourage the further development and clinical translation of FLT3-specific UniCAR T cells for the therapy of AML.
22 Feb 2023
TL;DR: In this paper , the authors investigate DNS resolution in a strict IPv6-only scenario and find that a substantial fraction of zones cannot be resolved, since the full DNS delegation chain must resolve via IPv6 as well.
Abstract: DNS is one of the core building blocks of the Internet. In this paper, we investigate DNS resolution in a strict IPv6-only scenario and find that a substantial fraction of zones cannot be resolved. We point out, that the presence of an AAAA resource record for a zone's nameserver does not necessarily imply that it is resolvable in an IPv6-only environment since the full DNS delegation chain must resolve via IPv6 as well. Hence, in an IPv6-only setting zones may experience an effect similar to what is commonly referred to as lame delegation. Our longitudinal study shows that the continuing centralization of the Internet has a large impact on IPv6 readiness, i.e., a small number of large DNS providers has, and still can, influence IPv6 readiness for a large number of zones. A single operator that enabled IPv6 DNS resolution -- by adding IPv6 glue records -- was responsible for around 20.3% of all zones in our dataset not resolving over IPv6 until January 2017. Even today, 10% of DNS operators are responsible for more than 97.5% of all zones that do not resolve using IPv6.
TL;DR: In this article , the authors proposed a switchable RevCAR system to target both the epidermal growth factor receptor (EGFR) and the disialoganglioside GD2, which are expressed in GBM.
Abstract: Glioblastoma (GBM) is still an incurable tumor that is associated with high recurrence rate and poor survival despite the current treatment regimes. With the urgent need for novel therapeutic strategies, immunotherapies, especially chimeric antigen receptor (CAR)-expressing T cells, represent a promising approach for specific and effective targeting of GBM. However, CAR T cells can be associated with serious side effects. To overcome such limitation, we applied our switchable RevCAR system to target both the epidermal growth factor receptor (EGFR) and the disialoganglioside GD2, which are expressed in GBM. The RevCAR system is a modular platform that enables controllability, improves safety, specificity and flexibility. Briefly, it consists of RevCAR T cells having a peptide epitope as extracellular domain, and a bispecific target module (RevTM). The RevTM acts as a switch key that recognizes the RevCAR epitope and the tumor-associated antigen, and thereby activating the RevCAR T cells to kill the tumor cells. However, in the absence of the RevTM, the RevCAR T cells are switched off. In this study, we show that the novel EGFR/GD2-specific RevTMs can selectively activate RevCAR T cells to kill GBM cells. Moreover, we show that gated targeting of GBM is possible with our Dual-RevCAR T cells, which have their internal activation and co-stimulatory domains separated into two receptors. Therefore, a full activation of Dual-RevCAR T cells can only be achieved when both receptors recognize EGFR and GD2 simultaneously via RevTMs, leading to a significant killing of GBM cells both in vitro and in vivo.