Advanced BioTech Solutions for Healthcare
Department of Pharmaceutical Sciences, Bouve College of Health Sciences
Vaccine Delivery Methods
Single Cell Studies
Vaccine Delivery Methods
Single Cell Studies
Vaccine Delivery Methods
Single Cell Studies
Vaccine Delivery Methods
The use of nano-liter reaction volumes and parallel sample processing offered by microfluidic devices make them ideally suited to total chemical and bioassay analysis, ultra-high throughput screening applications, and other cases where samples and reagents are available in limited quantities.
Our lab is focused on developing novel Bio-MEMS approaches to advance point of care diagnostics, cell culture and drug screening and delivery methods. We have developed Lab-on-a-Chip (LOC) devices that integrate several laboratory functions such as real time monitoring of target clinically relevant analyte, proteomics, genomics, cell-cell interactions as well as cell secretion and surface monitoring of single cells on a micro-chip. The advantages of the microfluidic based LOC developed by our lab include, the pico and nano-liter volumes, elimination of cross-contamination, the fast and efficient mixing of the reagents and gases and the ability to manipulate and analyze cells at a very high-throughput.
Our droplet based bio-assay platforms:
Cell-based therapy for modulating the immune response has gained momentum in recent years.
A promising approach to facilitate immune activation is the adoption of tissue engineering technology to generate three-dimensional (3D) constructs for cell function, cell-cell interaction studies, drug screening as well as cells and drugs delivery. We have developed a droplet microfluidic based technology to generate and monitor effects of therapeutic regimen on biomimetic micro-tumors tissue-chip. The microfluidic device is capable of generation and in-situ analysis of the micro-tumors for different applications in a high-throughput manner. It is equipped with a continuous perfusion module for continuous drug treatment and renewal of media and nutrients for the experimental duration to mimic in vivo conditions.
Furthermore, our laboratory is investigating a localized tissue engineered approach using 3D biocompatible materials to construct bio-matrixes loaded with vaccine and cells to enhance the immunologic milieu in patients. These constructs may serve as “lymphoid-like” constructs engineered to be implanted in vivo to deliver long-term immunity and specifically target cells to induce necessary mechanisms.
Cell migration and interactions in our bioengineered lymphoid-like constructs
The research program in our lab is focused on two general categories: 1. Development, evaluation and delivery of new therapeutic strategies and 2. Development of novel diagnostic and detection tools. In particular, we are working on a novel approach to develop and deliver cell-based therapy for modulating the immune response in cancer via bioengineered “lymphoid-like” constructs while our Lab-on-a-Chip (LOC) devices applied towards both analysis and manipulation of immunological system in cancer patients to overcome the disease.
Furthermore, we are applying our novel biotech methods to develop robust and portable device for detection of time-varying concentrations in real-time, of soluble molecules such as dosed insulin in T1D patients and phenotypic-based antimicrobial susceptibility testing (AST). Successful implementation of these research activities will represent a transformative advance in the field of diagnostics, therapy, biosensors, pharmacokinetic analysis and immunoassay development in microfluidic format.
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Single cell functional multi-omic analysis
Given the phenotypic and functional diversity of immune and cancer cell subsets, and the disparate timescale of immune reactions (early vs. delayed and transient vs. stable responses) it is essential to assess immune cell signaling and its target cell responses at single-cell level and more importantly in dynamic fashion to reveal the extent of heterogeneity. Our lab has developed h a microchip based method to evaluate cells and cellular interactions that can provide both high throughput and high content information bridging the gap between traditional assay plate in vitro methods and animal/human models.
Konry lab has developed an in-situ droplet microfluidic based technology to generate and monitor effects of therapeutic regimen on biomimetic micro-tumor tissues. The microfluidic device is capable of generation and in-situ analysis of the immunogenic micro-tumors for different applications in a high-throughput manner. It is equipped with a continuous perfusion module for continuous drug treatment and renewal of media and nutrients for the experimental duration to mimic in vivo conditions. Since 2016, our published in Lab on a chip journal breast tissue-chip model was numerously cited and serves as a basis for our current tissue-chip studies.
We have also extended our microfluidic approach to establish Diffuse Large B cell lymphoma (DLBCL). Diffuse large B cell lymphoma (DLBCL), the most common subtype of Non-Hodgkin lymphoma, exhibits pathologic heterogeneity and a dynamic immunogenic tumor microenvironment (TME). However, the lack of preclinical in vitro models of DLBCL TME hinders optimal therapeutic screening. Thus we have focused on the development of an integrated platform for high-throughput generation of immunogenic DLBCL models in a novel hydrogel combination of alginate and puramatrix, which promoted cell adhesion and aggregation. We demonstrated this system in dynamic analysis of cellular interaction, proliferation and therapeutic efficacy via spatiotemporal monitoring and secretome profiling. This work was recently accepted to Journal of Controlled Release (2019).
ScanDrop for rapid bacteria diagnostics and susceptibility testing
Urinary tract infections (UTIs) are among the most frequently encountered bacterial infections in the United States with an annual incidence over 8 million. In hospitalized patients, empiric therapy is given for 48-72 hours until traditional culture results and susceptibility data are available. However, the rapid emergence of antibiotic resistance presents an alarming challenge for management. It is now increasingly likely that many patients will be treated with inactive therapy, leading to adverse outcomes. Therefore, the development of new technologies to shrink the empiric therapy window through rapid identification and antibiotic susceptibility testing (AST) are critical. We have developed a novel technology called ScanDrop which incorporates a bead-based assay and microfluidics device to address shortcomings of current technology. As conceived, ScanDrop provides ultrafast, highly sensitive, direct-from-patient sample diagnostics for the most prevalent and antibiotic-resistant UTI pathogens (Escherichia coli, Klebsiella pneumoniae, Enterobacter spp., Pseudomonas aeruginosa, and Proteus spp.) without the need for culture pre-amplification. Our highly parallelized droplet microfluidic platform can screen four antibiotics/pathogens simultaneously and assess antibiotic sensitivity in 15-30 min. The advantages of our droplet-based AST, including rapid drug sensitivity response, morphological analysis and heterogeneity in antibiotic-resistance profiles, make it an excellent alternative to standard phenotypic AST with potential applications in clinical diagnostics and point of care testing.
Cloud-Enabled Microscopy and Droplet Microfluidic Platform for Specific Detection of Escherichia coli in Water
Here we have developed a ScanDrop platform combined of droplet microfluidics and portable imaging system which was integrated with cloud-based control software and data storage developed by our collaborator Dr.Hillman, UC Berkeley. The cloud-based control software and data storage enables robotic image acquisition, remote image processing, and rapid data sharing. These features form a “cloud” network for water quality monitoring. We have demonstrated the capability of ScanDrop to perform water quality monitoring via the detection of an indicator coliform bacterium, Escherichia coli, in drinking water contaminated with feces. Magnetic beads conjugated with antibodies to E. coli antigen were used to selectively capture and isolate specific bacteria from water samples. The bead-captured bacteria were co-encapsulated in pico-liter droplets with fluorescently-labeled anti-E. coli antibodies, and imaged with an automated custom designed fluorescence microscope. The entire water quality diagnostic process required 8 hours from sample collection to online-accessible results compared with 2-4 days for other currently available standard detection methods.
Bioartificial lymphoid organ for immune-regulation studies
One of most common cell therapy delivery methods is via systemic injection of the cells. This approach has several major drawbacks: 1) very large numbers of injected cells are required, and 2) most of the injected cells disappear within 24 hours post injection, and 3) side effects are common. To enhance the immunologic milieu to promote T cell activation our group in collaboration with Dr.Avigan is focused on a more localized tissue engineered approach using implantable three-dimensional (3D) scaffold constructs loaded with DC/tumor fusions cells. These constructs will serve a “lymphoid-like” organs engineered to be delivered in vivo to deliver DC/tumor fusions and specifically target T cells to induce anti-cancer mechanisms. We conducted the ex vivo studies to demonstrate that DC/tumor fusions can be seeded and maintained a live as well as allow the T cell migration into the scaffold and activation. Furthermore we there able to demonstrate in the scaffold that DC/fusions uniquely stimulate both helper and cytotoxic T cell responses through the presentation of internalized and newly synthesized antigens, respectively. This 3D ex vivo matrix microenvironment allowed us studying the cell-cell interaction of DC/fusions and T cells and predicate the vaccine effect in the in vivo.
Pico-liter droplet microfluidic immunosorbent platform for point-of-care diagnostics of tetanus
We have developed a sensitive, specific, rapid and low cost pico-liter microsphere based platform for bio-analyte detection and quantification. In this method, a biological sample, biosensing microspheres, and fluorescently labeled detection (secondary) antibodies are co-encapsulated to capture the analyte (here: human anti-tetanus immunoglobulin G) on the surface of the microsphere in microfluidic pL-sized droplets. The absorption of the analyte and detecting antibodies on the microsphere concentrate the florescent signal in correlation with analyte concentration. Using our platform and commercially available antibodies, we quantified the anti-tetanus antibodies content in human serum. In comparison to standard bulk immunosorbent assays, the microfluidic droplet platform presented here reduces the reagent volume by four orders of magnitude, while fast reagent mixing reduces the detection time from hours to minutes. We consider this platform to be a major leap forward in the miniaturization of immunosorbent assays and to provide a rapid and low cost global point-of-care tool.
Ultrasensitive Detection of Low-Abundance Surface- Marker Protein in a Microfluidic Nanoliter Platform
We have made significant headway in the development of droplet based microfluidic lab on chip Immuno-Rolling Circle Amplification (RCA) labeling assay with single-molecule sensitivity and ultrahigh detection specificity of antigen-antibody reactions to identify specific cell surface markers . To take full advantage of the RCA labeling protocol for identifying low abundance proteins, we applied the RCA reaction inside monodisperse aqueous emulsion nano-liter droplets containing live single cells. By merging the single molecule detection power of the RCA strategy with microfluidic technology, we were able to demonstrate that identification of specific tumor marker, EpCAM, can be achieved on tumor-cell surface in miniaturized nanoliter reaction droplets.
Approaching near real-time biosensing: microfluidic microsphere based biosensor for real-time analyte detection.
The objectives of this project is to develop a novel immunoassay-based microfluidic sensor to assess multiplexed detection of physiological markers. The proposed platform is in line with the long-sought goal in modern medicine to develop smart sensors that monitor physiological conditions in real-time and deliver therapeutics in a localized, controllable manner. For that aim we have developed a MATLAB tracking/detection algorithm which detects moving microspheres in each recorded frame in a detection channel. We have already initiated prototyping the microsphere tracking and detection software in the proposed LOC as can be seen in the recorded movie bellow. Such sensing networks can improve healthcare dramatically by personalizing diagnostics for better clinical outcome and providing real-time feedback on sensing and actuation.
Development of continuous interstitial insulin monitoring approach in T1D for optimizing the performance of bionic pancreas systems
Type 1 Diabetes (T1D) is a chronic disease with no cure as of 2011. Even with state-of-the-art insulin therapy using rapid-acting insulins, substantial hyperglycemia and hypoglycemia persist in most people with T1D, in part because of the difficulties of calculating the appropriate insulin dose for a given situation, and in part due to the discontinuous nature of therapy.
Development of an approach to monitor interstitial insulin in real-time would allow the pharmacokinetic (PK) characteristics of insulin analogs to be determined in real-time. Currently no self-monitoring approach is available for continuous insulin measurements and PK characteristics of insulin in real-time. We are working together in collaboration with Dr. Steven Russell, MGH Diabetes Unit on the development of new microfluidic lab-on-a-chip sensor device that continuously samples the subcutaneous interstitial fluid using microdialysis and then measures insulin levels in the microdialysate continuously (Fig.1C). A polydimethylsiloxane (PDMS) based microfluidic platform (Fig.1B) that involves a microsphere-based immunometric insulin detection scheme is currently being developed (Fig.1A). Once developed, the device can be integrated into a next generation bionic pancreas system which will then adjust parameters in its insulin PK model according to data received from the continuous insulin monitor. We can then test the hypothesis that real-time insulin monitoring and PK analysis will improve the performance of the bionic pancreas system.
Phenotypic drug profiling in droplet microfluidics for better targeting of drug-resistant tumors.
Screening known chemotherapeutic drugs and novel chemical reagents against cancer cells is necessary for the pharmaceutical industry and identification of drug targets. At present, this time consuming, laborious process is performed in large batches of micro-titre plates. We are developing droplet-based single cell drug profiling platforms that allow us to monitor hundreds of individual reactions, quantify cell responses and correlate drug cytotoxicity in breast cancer cells. Our aim is to develop large-scale multiparametric cytological profiles of cells to characterize dose-dependent therapeutic response in the treatment of multi-drug resistant blood and breast cancers.
Live single cell functional phenotyping and cell-cell communication in droplet nano-liter reactors
The immune system plays an integral role in host defense, cancer progression, and inflammation. Substantial progress in understanding mechanisms of immune regulation in tumors, allergy, asthma, autoimmune diseases, organ transplantation and chronic infections will lead to a variety of possibilities for targeted therapeutic approaches. Our research is focused on the application of bioengineering principals to both analysis and manipulated immunological system. In particular our research program involves in a single-cell molecular phenotype characterization and cell-cell interactions that addresses fundamental biological questions and allows one to observe functional proteomic phenomena in heterogeneous populations of single cells. This project outlines the integration of a novel microfluidic approach developed by us to monitor live cell surface expression changes as well as the actual immunological synapse (IS) (R21/NIH/NCI, funded 2012-2014, PI T.Konry). In addition to cell surface studies, this approach will also allow us to collect data on secreted molecules during IS formation. Thus, live cell secretion and cell surface changes will be monitored on a single cell level simultaneously in a distinct microenvironment, feat not possible using conventional techniques. Such approach would have a big impact on understanding biological processes, which are inherently dynamic.
Dr. Alex Lim, Postdoctoral Fellow
Dr. Stefano Ugolini, Postdoctoral Fellow
Ilana Berger Fridman, PhD student
Sullivan Matt, PhD student
Jose Estevam, Grad student
Yuntong Wang, Zhi Lin Shen, Millicent Gabriel, Awdhoot Godbole, Radhika Kulkarni, Ria Rajesh Bedi, Xinyi Shao, Suchitra Ramesh, Supriya Nagarajan, Viren Bhatia
Michael Gray, Amey Gaikwad, Dr.Saheli Sarkar, Wenjing Kang, Seamus McKenney, Kristy Fang, Sai Mynampati, Abhishek Chiyyeadu,Sayalee Potdar,Sneha Pawar,Kristy Fang,Rucha Adhav,Chanchal Rathi,Himali Shroff,Chaitra Belgur, Dipen Parande, Vinny Motwani, Abhinav Gupta, Sneha Varghese, Dishant Patel, Harnil Shah, Noa Cohen, Micah AmdurClark, Pooja Sabhachandani, Dr. Yantao Fan
Dr. Konry was awarded with CTSI Tufts and RADX/NIH Phase 0 grants for her PCR free RAPID testing technology for COVID-19 diagnostics
Collaboration in immunotherapy between Konry Lab NEU and Dr.R.Romee Dana Farber
Drs.Konry and Romee were awarder with a grant from DFCI/NEU joined program in cancer drug development (2019-2021). The grant is focused on applying novel microfluidic technology to develop memory NK cell based therapy.
Dr.Konry was appointed to Women in Microfluidics & BioMEMS list.
Casis tissue NASA
Konry lab was awarded a collaborative NASA tissue grant to study the effect of the extreme environment of space on immune cell interactions.
Dr.Konry was awarded with 2017 Tufts Clinical and Translational Science Institute (CTSI) Pilot Award
The project will focus on determining antibody treatment sensitivity in B cell lymphoma by novel microfluidics-based NK cell immunogenicity platform developed by Dr.Konry’s group (Collaboration with Dr. Andrew Evens , Tufts Medical Center).
Pooja Sabhachandani will present at SLAS 2017 conference our work on novel droplet based platform for biomimetic tumor microenvironment studies and conduct the SLAS 2017 – Podium Presentation Webinar
The project is focused on developing a device for rapid urinary tract infection diagnosis and antibiotic susceptibility testing.
Our new findings in dynamic analysis of immune and cancer cell interactions: a single cell lab on a chip perspective
We investigated the dynamics of live cell anti-tumor immune responses at the single cell level in a microfluidic platform. The integrated droplet array allowed improved control over heterotypic cell pairing and interactions, which allowed us to observe significant cell motility, morphological changes, and complex formation over an extended duration. We evaluated immune cell priming by Ag-loaded DCs and the subsequent functional outcome in the con- text of multiple myeloma cells. Our results demonstrate substantial heterogeneity in priming interactions between DC and T cells, both in basal and activated cells. Effector T cells depicted time-varying cytotoxicity following transient, short or long stable contacts. Serial interactions by T cells were observed both in upstream (DC-based) and downstream (target-based) interactions. Our future aims include determining the molecular mechanisms underlying the phenotypic heterogeneity in T cell responses in droplets, and integrated live cell analysis of immune cell activation and effector functions.
Our novel droplet‐merging platform has been applied for comparative functional analysis of M1 and M2 macrophages for Forsyth institute
In our recent paper published in Biotechnology and Bioengineering we presented a simple and effective method for the co-encapsulation of polarized M1 and M2 macrophages with Escherichia coli (E. coli) by passive merging in an integrated droplet generation, merging, and docking platform. This approach facilitated live cell profiling of effector immune functions in situ and quantitative functional analysis of macrophage heterogeneity.
Our recent publication in Lab on a chip:
Acquired drug resistance is a key factor in the failure of chemotherapy. Due to intratumoral heterogeneity, cancer cells depict variations in intracellular drug uptake and efflux at the single cell level, which may not be detectable in bulk assays. In this study we present a droplet microfluidics-based approach to assess the dynamics of drug uptake, efflux and cytotoxicity in drug-sensitive and drug-resistant breast cancer cells. An integrated droplet generation and docking microarray was utilized to encapsulate single cells as well as homotypic cell aggregates. Drug-sensitive cells showed greater death in the presence or absence of Doxorubicin (Dox) compared to the drug-resistant cells. We observed heterogeneous Dox uptake in individual drug-sensitive cells while the drug-resistant cells showed uniformly low uptake and retention. Dox-resistant cells were classified into distinct subsets based on their efflux properties. Cells that showed longer retention of extracellular reagents also demonstrated maximal death. We further observed homotypic fusion of both cell types in droplets, which resulted in increased cell survival in the presence of high doses of Dox. Our results establish the applicability of this microfluidic platform for quantitative drug screening in single cells and multicellular interactions.
Dr. Konry was awarded with Schumacher Faculty Award 2015, presented to one faculty member early in their Northeastern career to recognize significant academic achievement for work done at Northeastern University
Dr. Konry was awarded with competitive grant from DFCI/NU Joint Program in Cancer Drug Development in collaboration with Prof. Suzanne Gaudet (DF/HMS).
Congratulations to Pooja Sabhachandani on being awarded with Shevell/Cohen Cancer Research Award (first place winner)
Our recent publication in Biosensors and Bioelectronics:
Approaching near real-time biosensing: Microfluidic microspherebased biosensor for real-time analyte detection
In this study we describe a simple lab-on-a-chip (LOC) biosensor approach utilizing well mixed micro- fluidic device and a microsphere-based assay capable of performing near real-time diagnostics of clinically relevant analytes such cytokines and antibodies. We were able to overcome the adsorption kinetics reaction rate-limiting mechanism, which is diffusion-controlled in standard immunoassays, by introducing the microsphere-based assay into well-mixed yet simple microfluidic device with turbulent flow profiles in the reaction regions. The integrated microsphere-based LOC device performs dynamic detection of the analyte in minimal amount of biological specimen by continuously sampling micro-liter volumes of sample per minute to detect dynamic changes in target analyte concentration. Furthermore we developed a mathematical model for the well-mixed reaction to describe the near real time detection mechanism observed in the developed LOC method. To demonstrate the specificity and sensitivity of the developed real time monitoring LOC approach, we applied the device for clinically relevant analytes: Tumor Necrosis Factor (TNF)alpha cytokine and its clinically used inhibitor, anti-TNF-α antibody. Based on the reported results herein, the developed LOC device provides continuous sensitive and specific near real-time monitoring method for analytes such as cytokines and antibodies, reduces reagent volumes by nearly three orders of magnitude as well as eliminates the washing steps required by standard immunoassays.
Our recent publication in PlosOne:
Cloud-Enabled Microscopy and Droplet Microfluidic Platform for Specific Detection of Escherichia coli in Water
Published: January 27, 2014 DOI: 10.1371/journal.pone.0086341
We report an all-in-one platform – ScanDrop – for the rapid and specific capture, detection, and identification of bacteria in drinking water. The ScanDrop platform integrates droplet microfluidics, a portable imaging system, and cloud-based control software and data storage. The cloud-based control software and data storage enables robotic image acquisition, remote image processing, and rapid data sharing. These features form a “cloud” network for water quality monitoring. We have demonstrated the capability of ScanDrop to perform water quality monitoring via the detection of an indicator coliform bacterium, Escherichia coli, in drinking water contaminated with feces. Magnetic beads conjugated with antibodies to E. coli antigen were used to selectively capture and isolate specific bacteria from water samples. The bead-captured bacteria were co-encapsulated in pico-liter droplets with fluorescently-labeled anti-E. coli antibodies, and imaged with an automated custom designed fluorescence microscope. The entire water quality diagnostic process required 8 hours from sample collection to online-accessible results compared with 2-4 days for other currently available standard detection methods.
Our lab in the news
DOE Science Highlights: A cool glass of clean drinking water. A new system called ScanDrop could revolutionize how we identify pathogens in drinking water
Tania (Tali) Konry, Ph.D. Associate Professor Department of Pharmaceutical Sciences Northeastern University 140 The Fenway, R 441, Lab 446 Boston, MA 02115 Tel: 617.373.3224