Publications

Personalised cell therapies utilising T cell receptors TCRs show tremendous clinical promise though TCR synthesis and validation techniques lag far behind current TCR repertoire sequencing capacity To address this gap we developed makeTCR a modular TCR cloning system that enables rapid single-step fidelity assembly of human or murine TCR sequences ...More |Related Solutions: Mantis^®

Personalised cell therapies utilising T cell receptors (TCRs) show tremendous clinical promise, though TCR synthesis and validation techniques lag far behind current TCR repertoire sequencing capacity. To address this gap we developed makeTCR: a modular TCR cloning system that enables rapid, single-step, 100% fidelity assembly of human or murine TCR sequences into diverse expression vectors. We provide pre-cloned modules for αβ and γδ TCRs, as well as many native and engineered constant regions. We show how implementing cell-free manufacturing both facilitates the propagation of precloned modules, and allows testable TCR material to be synthesised in 24 hours, enabling patient-derived TCRs to be prototyped prior to use in personalised cell therapies. makeTCR scales to making thousands of TCRs, at high fidelity and at substantially reduced cost. makeTCR is facilitated by a free, open-source, extensible, graphical platform to simplify, standardise, and accelerate TCR functionality testing for personalised medicine and beyond. Less |Related Solutions: Mantis^®

Background Selective and potent Toll-like receptor TLR agonists are currently under evaluation in preclinical models and clinical studies to understand how the innate immune system can be harnessed for therapeutic potential These molecules are designed to modulate innate and adaptive immune responses making them promising therapeutic candidates for treating diseases ...More |Related Solutions: Mantis^®

Background: Selective and potent Toll-like receptor (TLR) agonists are currently under evaluation in preclinical models and clinical studies to understand how the innate immune system can be harnessed for therapeutic potential. These molecules are designed to modulate innate and adaptive immune responses, making them promising therapeutic candidates for treating diseases such as cancer or chronic viral infections. Much is known about the expression and signaling of TLRs which varies based on cell type, cellular localization, and tissue distribution. However, the downstream effects of different TLR agonists on cellular populations and phenotypes are not well understood. This study aimed to investigate the impact of TLR pathway stimulation on peripheral blood mononuclear cell (PBMC) cultures from people living with HIV (PLWH) and healthy donors.

Methods: The effects of TLR4, TLR7, TLR7/8, TLR8 and TLR9 agonists were evaluated on cytokine production, cell population frequencies, and morphological characteristics of PBMC cultures over time. Changes in the proportions of different cell populations in blood and morphological features were assessed using high-content imaging and analyzed using an AI-driven approach.

Results: TLR4 and TLR8 agonists promoted a compositional shift and accumulation of small round (lymphocyte-like) PBMCs, whereas TLR9 agonists led to an accumulation of large round (myeloid-like) PBMCs. A related increase was observed in markers of cell death, most prominently with TLR4 and TLR8 agonists. All TLR agonists were shown to promote some features associated with cellular migration. Furthermore, a comparison of TLR agonist responses in healthy and HIV-positive PBMCs revealed pronounced differences in cytokine/chemokine responses and morphological cellular features. Most notably, higher actin contraction and nuclear fragmentation was observed in response to TLR4, TLR7, TLR7/8 and TLR9 agonists for antiretroviral therapy (ART)-suppressed PLWH versus healthy PBMCs.

Conclusions: These data suggest that machine learning, combined with cell imaging and cytokine quantification, can be used to better understand the cytological and soluble immune responses following treatments with immunomodulatory agents in vitro. In addition, comparisons of these responses between disease states are possible with the appropriate patient samples. Less |Related Solutions: Mantis^®

Insulin is a key hormone in glucose homeostasis Its lack causes severe health complications and has to be compensated by regular administration of insulin Despite intense long-lasting research a more stable substitute has yet to be discovered to alleviate patients' issues Here we report the development of a novel assay ...More |Related Solutions: Mantis^®

Insulin is a key hormone in glucose homeostasis. Its lack causes severe health complications and has to be compensated by regular administration of insulin. Despite intense long-lasting research, a more stable substitute has yet to be discovered to alleviate patients' issues. Here we report the development of a novel assay for screening potential insulin analogues based on the recently published method DIANA. Our assay meets the need for a fast, non-radioactive method as a sensitive alternative to the commonly used radioactive immunoassay. Less |Related Solutions: Mantis^®

Cells of the myeloid lineage particularly monocytes and macrophages play a key role in HIV infection by contributing to viral replication immune response and maintaining immune balance during suppressive therapy We hypothesized that metabolic reprogramming and altered chemokine signaling in people living with HIV PWH on long-term antiretroviral therapy ART ...More |Related Solutions: Mantis^®

Cells of the myeloid lineage, particularly monocytes and macrophages, play a key role in HIV infection by contributing to viral replication, immune response, and maintaining immune balance during suppressive therapy. We hypothesized that metabolic reprogramming and altered chemokine signaling in people living with HIV (PWH) on long-term antiretroviral therapy (ART) affect monocyte transport and polarization due to ongoing inflammation. Therefore, the present study aimed to identify the mechanism of impaired monocyte/macrophage function in PWH on well-treated ART that can lead to clinical intervention strategies to improve health. Single-cell RNA sequencing, immune-phenotyping, and metabolic modeling identified altered expression of chemokine and metabolite receptors and altered metabolic flux in PWH monocytes that decreased monocyte migration. The plasma secretome revealed a nonclassical inflammatory microenvironment in PWH. Integrative multi-omics and single-cell proteomics of differentiated monocyte-derived macrophages (MDMs) detected metabolic reprogramming orchestrated by α-ketoglutarate (AKG) that affected macrophage function and HIV infection. Increased levels of AKG in plasma were shown to occur in PWH under ART. Therefore, when differentiating MDM with serum from PWH or AKG, macrophage function was found polarized towards an M2-like state. AKG alone was shown to increase CCR5 levels and increase HIV-1 infection in MDM. Here, we utilize systems biology-driven identification and ex vivo assays to show impaired macrophage polarization, due to metabolic training, can leads to a low-grade nonclassical inflammatory environment in well-treated PWH. Less |Related Solutions: Mantis^®

High-grade serous ovarian cancer HGSOC is a devastating disease that is frequently detected at an advanced and incurable stage Advances in ultrasensitive mass spectrometry-based spatial proteomics have provided a unique opportunity to uncover early molecular events in tumorigenesis and common dysregulated pathways with high therapeutic potential Here we present a ...More |Related Solutions: Mantis^®

High-grade serous ovarian cancer (HGSOC) is a devastating disease that is frequently detected at an advanced and incurable stage. Advances in ultrasensitive mass spectrometry-based spatial proteomics have provided a unique opportunity to uncover early molecular events in tumorigenesis and common dysregulated pathways with high therapeutic potential. Here, we present a comprehensive proteomic analysis of serous tubal intraepithelial carcinoma (STIC), the HGSOC precursor lesion, covering more than 10,000 proteins from ultralow input archival tissue. We discovered that STICs and concurrent invasive carcinomas were indistinguishable at the global proteome level, revealing a similar level of phenotypic and molecular heterogeneity. Using cell-type resolved tissue proteomics, we revealed strong cell-of-origin signatures preserved in STICs and invasive tumors and identified early dysregulated pathways of therapeutic relevance. These include proliferation and DNA damage repair signatures, as well as onco-metabolic changes, such as increased cholesterol biosynthesis. Finally, we uncovered substantial remodeling of the co-evolving tumor microenvironment, affecting approximately one-third of the stromal proteome, and derived a common signature associated with progressive immunosuppression and ECM restructuring. In summary, our study highlights the power of spatially resolved quantitative proteomics to dissect the molecular underpinnings of early carcinogenesis and provides a rich proteomic resource for future biomarker and drug target discovery. Less |Related Solutions: Mantis^®