Biology subjects

Skardal, A.

Publications and source records attributed to Skardal, A..

8 recordsLinked to original sources

bioRxiv2026

Development of a Thyroid Cancer to Lung Metastasis-on-a-Chip Model

Background: Study of the metastatic cascade in differentiated non-medullary thyroid cancer (NMTC) is hampered by the relative lack of preclinical models that metastasize from primary tumors to other organs. We previously identified regulator of calcineurin 1 isoform 4 (RCAN1.4) as a suppressor of NMTC cell growth, invasion, and metastasis using in vitro systems and tail vein injections in immunocompromised mice. We then reported a microfluidic device mimicking the mouse tail vein injection where cancer cells were allowed to flow directly to lung organoid constructs. We now report the development of a full primary tumor to lung metastasis-on-a-chip (MOC) model for TC. Methods: Resin molds for primary and secondary sites connected by a channel were 3D printed and filled with polydimethylsiloxane to create a molded silicone elastomer. The primary site was seeded with an organoid composed of a hyaluronic acid and collagen-based hydrogel into which FTC236 and HTh74 NMTC cells with and without knockdown of RCAN1.4 were engrafted. The secondary site consisted of a hydrogel organoid into which bronchial epithelial cells and lung fibroblasts were seeded. Culture media was circulated and movement of NMTC cells to the secondary site was assessed. Results: Both cell lines that expressed RCAN 1.4 metastasized to the secondary lung site. RCAN1.4-reduced FTC236 cells had increased metastatic behavior vs controls while HTh74 was less affected by RCAN1.4 loss, similar to prior studies. To determine if the MOC could be used for incorporation of immune components and therapeutic modeling, NK-92 cells were added and were shown to be reduce metastases for both cell lines in the presence or absence of RCAN1.4. Analysis of MOC circulating fluid revealed elevated levels of granzyme B, consistent with NK-92 cell activation. Cytokine profiling identified elevated probable NMTC and NK-92-derived factors. Conclusions: These results indicate that NMTC metastatic behavior can be modeled using a novel MOC system for mechanistic and preclinical therapeutic studies.

cancer biology↗

bioRxiv2026

Engineering a 3D Lung Co-culture Platform to Model Epithelial-Fibroblast Interactions in Pulmonary Fibrosis

Idiopathic pulmonary fibrosis (IPF) is a fatal interstitial lung disease (ILD) characterized by progressive fibrosis, irreversible loss of lung elasticity, and chronic respiratory failure, with a mean survival of 3-5 years. The disease is believed to result from repeated alveolar epithelial injury that sustains transforming growth factor-beta (TGF-{beta}) signaling, driving fibroblast-to-myofibroblast differentiation and excessive collagen deposition. Although current IPF models--including animal studies, 2D cultures, and basic 3D systems--have enhanced understanding of disease mechanisms, they inadequately replicate epithelial-fibroblast interactions, extracellular matrix (ECM) remodeling, and epithelial barrier dysfunction. To address this limitation, we engineered a 3D lung co-culture model that simulates the physiological epithelial-fibroblast crosstalk and ECM remodeling characteristic of IPF. Our model embeds fibroblasts within a collagen-hyaluronic acid matrix overlaid with an epithelial monolayer cultured at an air-liquid interface. Basolateral TGF-{beta} exposure generated a profibrotic microenvironment that weakened epithelial barrier integrity and drove myofibroblast differentiation marked by elevated -SMA and vimentin. Elevated pro-inflammatory cytokine secretion and increased collagen disorganization further demonstrated active fibrogenesis. Together, these features show that our model captures key early events in IPF pathogenesis and offers a versatile platform for next-generation lung-on-a-chip studies in fibrotic disease.

bioengineering↗

bioRxiv2026

Establishment of a patient-derived adrenocortical carcinoma 3D tumor construct platform for evaluation of therapeutic strategies

Adrenocortical carcinoma (ACC) is an under-studied, aggressive cancer of the adrenal glands where surgical resection is currently the only effective curative option. However, after surgery the majority of patients experience tumor progression. The median overall survival of 12 months has not improved since approval of mitotane in 1970. The lack of effective therapies in ACC is partially due to the lack of preclinical models that accurately represent human ACC. Most attempts to generate ACC cell lines or animal models have been unsuccessful. The few existing models do not adequately reflect the oncogenic signaling pathways or intratumoral heterogeneity of human ACC. These limited model systems have hindered identification of drivers of tumor progression and immune escape mechanisms, thereby limiting development and testing of novel therapeutic approaches such as targeted therapies or immunotherapies. We developed human ACC patient-derived tumor constructs (PTCs) encapsulated in synthetic extracellular matrix. Our ACC PTCs exhibit hallmarks of ACC: proliferation, expression of key ACC biomarkers, such as SF1, and production of cortisol. We provide characterization in the form of immunofluorescence staining for ACC biomarkers, and confirmation of PTC proliferation and cortisol production - a hallmark of ACC. We then demonstrate the utility of ACC PTCs for evaluation of chemotherapies currently used clinically (mitotane) with and without experimental cocktails of etoposide, doxorubicin, and cisplatin, experimental targeted therapies, and cellular immunotherapies, primarily in the form of natural killer (NK) cell therapy. In particular, the latter - cellular immunotherapies - are cutting edge studies demonstrating potential to evaluate immunotherapies in ACC clinical scenarios. Together these data provide evidence that patient-derived ACC models can serve as an important tool for identification of future points of intervention and testing of novel therapeutic strategies to improve ACC clinical care.

cancer biology↗

bioRxiv2025

Evaluation of natural killer cell tumor homing and effector function in response to CDK4/6 and AURKA inhibition in a melanoma tumor-on-a-chip platform

Natural killer (NK) cells have emerged as an important clinical tool cellular immunotherapy. Whereas immune checkpoint blockade (ICB) or chimeric antigen receptor (CAR) T-cell therapy (CAR-T) therapy have been adopted as a first line treatments in different malignancies, such as melanoma, these approaches do not work for all patients. T cells require proper antigen presentation on tumor cells for recognition and to carry out their corresponding cytotoxic functions. Deficiency of tumor antigens, or high variability in those present, make T cell-based CAR-T and ICB ineffective. By contrast, NK cells are not limited by antigen presentation deficiencies, offering a potential alternative approach, yet their efficacy can suffer from immunosuppressive signals. Herein, we sought to develop in vitro and on-chip platforms to identify strategies for enhance, rather than suppress, NK cell homing to tumor cells. We explored the use of inhibition of kinases such as CK4/6 and AURKA to induce tumor cell production of chemokines that NK cells migrate towards in aggressive melanoma models. We evaluated chemokine-aided NK cell migration-homing capabilities and their therapeutic efficacy and found that treatment of both melanoma cell line and patient-tumor constructs (PTCs) with CDK4/6 and AURKA generally resulted in improved NK cell homing to tumor cells and accompanying tumor cell killing. Interestingly, this chemokine-guided NK cell migration did not generate as effective outcomes in models using a mildly aggressive melanoma cell line. For our studies, we used 3D tumor constructs in both static Transwell models and then in a bioengineered NK cell-functionalized tumor-on-a-chip (NK-TOC) platform. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=83 SRC="FIGDIR/small/662476v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@17bf445org.highwire.dtl.DTLVardef@e1e1d9org.highwire.dtl.DTLVardef@1b29c2corg.highwire.dtl.DTLVardef@12b1f03_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2025

Fibroblasts drive differential response in immunotherapy via IL-6 in bioengineered 3D cancer constructs

Despite various therapeutic advancements in cancer, heterogeneity in the tumor microenvironment is a major hindrance to the overall efficacy of treatment. In this study, we focus on the most abundantly found cellular stromal components - fibroblasts - which can be tumor-promoting or tumor-suppressing based on the extent of disease progression and numerous additional factors. In the studies described herein, we evaluated their role in influencing chemotherapy as well as in immunotherapy using colorectal cancer (CRC) cell line-derived 3D tumor constructs. Our initial efforts centered on common chemotherapeutic agents utilized in CRC. However, while the presence of fibroblasts often did influence chemotherapy efficacy, a clear and consistent increase or decrease in efficacy was not obvious. We then focused subsequent studies on natural killer (NK) cell-based immunotherapy efficacy. There are numerous components and pathways involved in tumorigenesis and tumor-immune cell interactions, and the crosstalk between fibroblasts and immune cells like NK cells is still not well-defined. Fibroblasts have been documented to be a major player with respect to immune modulation and understanding this in greater detail can elucidate more options for anti-tumor immunity. Specifically, we used an extracellular matrix-based hydrogel platform to generate 3D CRC constructs to evaluate how fibroblasts affect tumor cell viability in presence and absence of NK cells. In general, fibroblasts aid the tumor cells in our models. We then identify a cytokine-based pathway (interleukin 6 [IL-6]) via which activated fibroblasts engage with the tumor cells and show how NK cell effector function can be restored when the IL-6 pathway is blocked in a melanoma-based tumor-on-a-chip platform. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=118 SRC="FIGDIR/small/660473v1_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@53797forg.highwire.dtl.DTLVardef@4dbc83org.highwire.dtl.DTLVardef@19346deorg.highwire.dtl.DTLVardef@11d834_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2023

A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity

Adrenocortical carcinoma (ACC) has a poor prognosis, and no new drugs have been identified in decades. The absence of drug development can partly be attributed to a lack of preclinical models. Both animal models and 2D cell cultures of ACC fail to accurately mimic the disease, as animal physiology is inherently different than humans, and 2D cultures fail to represent the crucial 3D architecture. Organoids and other small 3D in vitro models of tissues or tumors can model certain complexities of human in vivo biology; however, this technology has largely yet to be applied to ACC. In this study, we describe the generation of 3D tumor constructs from an established ACC cell line, NCI-H295R. NCI-H295R cells were encapsulated to generate 3D ACC constructs. Tumor constructs were assessed for biomarker expression, viability, proliferation, and cortisol production. In addition, matrix metalloproteinase (MMP) functionality was assessed directly using fluorogenic MMP-sensitive biosensors and through infusion of NCI-H295R cells into a metastasis-on-a-chip microfluidic device platform. ACC tumor constructs showed expression of biomarkers associated with ACC, including SF-1, Melan A, and inhibin . Treatment of ACC tumor constructs with chemotherapeutics demonstrated decreased drug sensitivity compared to 2D cell culture. Since most tumor cells migrate through tissue using MMPs to break down extracellular matrix, we validated the utility of ACC tumor constructs by integrating fluorogenic MMP-sensitive peptide biosensors within the tumor constructs. Lastly, in our metastasis-on-a-chip device, NCI-H295R cells successfully engrafted in a downstream lung cell line-based construct, but invasion distance into the lung construct was decreased by MMP inhibition. These studies, which would not be possible using 2D cell cultures, demonstrated that NCI-H295R cells secreted active MMPs that are used for invasion in 3D. This work represents the first evidence of a 3D tumor constructs platform for ACC that can be deployed for future mechanistic studies as well as development of new targets for intervention and therapies. SignificanceThe paucity of preclinical research models has contributed to lack of progress in treatment of adrenocortical carcinoma (ACC). Three-dimensional modeling of ACC may provide novel insights for preclinical studies and advance ACC research.

cancer biology↗

bioRxiv2021

Engineering a thixotropic and biochemically tunable hyaluronan and collagen bioink for biofabrication of multiple tissue construct types

The field of three-dimensional (3D) bioprinting has advanced rapidly in recent years. Significant reduction in the costs associated with obtaining functional 3D bioprinting hardware platforms is both a cause and a result of these advances. As such, there are more laboratories than ever integrating bioprinting methodologies into their research. However, there is a lack of standards in the field of biofabrication governing any requirements or characteristics to support cross-compatibility with biomaterial bioinks, hardware, and different tissue types. Here we describe a modular extracellular matrix (ECM) inspired bioink comprised of collagen and hyaluronic acid base components that: 1) employ reversible internal hydrogen bonding forces to generate thixotropic materials that dynamically reduce their elastic moduli in response to increased shear stress, thus enabling increased compatibility with printing hardware; and 2) modular addons in the form of chemically-modified fibronectin and laminin that when covalently bound within the bioink support a variety of tissue types, including liver, neural, muscle, pancreatic islet, and adipose tissue. These features aim to accelerate the deployment of such bioinks for tissue engineering of functional constructs in the hands of various end users.

bioengineering↗

bioRxiv2019

Simulating the human tumor microenvironment in colorectal cancer organoids in vitro and in vivo

The tumor microenvironment (TME) plays a significant role in cancer growth and metastasis. Bioengineered models of the TME will advance our understanding of cancer progression and facilitate identification of novel anti-cancer therapeutics that target TME components such as extracellular matrix (ECM) and stromal cells. However, most current in vitro models fail to recapitulate the extensive features of the human tumor stroma, especially ECM architecture, and are not exposed to intact body physiology. On the other hand, in vivo animal models do not accurately capture human tumor architecture. Using the features of biopsied colorectal cancer (CRC) tissue as a guide, we address these deficiencies by creating human organoids containing a colonic stromal ECM layer and CRC spheroids. Organoids were studied in vitro and upon implantation in mice for 28 days. We show that the stromal ECM micro-architecture, generated in vitro, was maintained in vivo for at least 28 days. Furthermore, comparisons with biopsied CRC tumors revealed that organoids with orderly structured TMEs induce an epithelial phenotype in CRC cells, similar to low-grade tumors, compared to a mesenchymal phenotype observed in disordered TMEs, similar to high-grade tumors. Altogether, these results are the first demonstration of replicating the human tumor ECM architecture in biofabricated tumor organoids under ex vivo and in vivo conditions.

cancer biology↗