The Reineke group specializes in the design and discovery of functional macromolecules for biomedical research

Synthetic materials are playing an important role in the diagnosis and treatment of many diseases. Consequently, understanding how novel macromolecules interact with and affect living systems is one of the most fundamental and important problems in biomedical research. The Reineke Research Group seeks to discover novel nucleic acid drugs, delivery vehicles, and imaging agents to diagnose and treat disease. Likewise we aim to elucidate the cellular mechanisms of biomaterial internalization, their function and elimination, and toxicity by designing and synthesizing novel systems to aid our understanding of these phenomena.

Our research efforts are interdisciplinary in nature and encompass the areas of chemistry, materials science, biology and medicine. Students in the Reineke Group become familiar with organic, inorganic, and polymer synthesis techniques and physical/analytical characterization procedures such as gel permeation chromatography, static and dynamic light scattering, multi-dimensional NMR, ¹H inversion recovery experiments, transmission electron microscopy, FT-IR, and mass spectrometry. In addition, students routinely gain expertise in gel electrophoresis, confocal microscopy, cell culture, and MRI methodologies. Our current research interests are listed below:

Carbohydrate-Containing Polymers and Click Clusters for Nucleic Acid Delivery

The wealth of information being obtained from genomic, proteomic, and glycomic research is allowing researchers to unravel the intricate genetic and epigenetic mechanisms associated with human health and disease. Ubiquitous tools such as miRNA (microRNA), siRNA (small interfering RNA), oligodeoxynucleotide (ODN) transcription factor decoys, plasmid DNA, aptamers, genetic vaccines, and many other polynucleotide forms are transforming the methods of regulating gene expression and epigenetic mechanisms for understanding biological processes, disease pathways, and are undergoing extensive research and development as novel therapeutics. Nucleic acids have exceptional affinity and specificity for their intracellular targets; yet, many complex factors dictate the accuracy, reproducibility, and relevance of utilizing polynucleotides as novel therapeutics. Indeed, the nucleic acid delivery vehicle plays a central yet elusive role in dictating the efficacy, safety, mechanisms, and kinetics of gene regulation in a spatial and temporal manner. Delivery systems are needed to compact nucleic acids into nanostructures, termed polyplexes, that can enter cells, protect nucleic acids from enzymatic damage, and provide the possibility of targeting the delivery to specific tissue types and sites within the cell. We strive to creatively design innovative polymeric vehicles for delivering gene regulatory nucleic acids and develop new experimental methods to gain a fundamental understanding of their interactions and pathways taken within living systems in a spatial and temporal manner. We have developed several novel carbohydrate-containing polymers that have shown outstanding affinity to form polyplexes and facilitate intracellular nucleic acid delivery efficiency with low toxicity. We are concerned with the design, synthesis, and biological characterization as well as with the examination of the mechanism of toxicity and gene delivery of polymers and click  clusters in cell culture.

Three main subprojects in this area focus on:

1. Elucidating how differences in the chemical structures of polymers and click clusters affect toxicity, efficiency, and mechanisms of DNA delivery.
2. Tissue specific targeting of novel nucleic acid therapeutics.
3. The delivery of DNA decoys to block NF-kappaB in heart tissue, a transcription factor known to mediate cardiovascular disease.

Glycopolymer-Gd³⁺ Chelates as Novel MRI Contrast Agents

The early diagnosis of devastating ailments such as heart disease and cancer plays a large role in the successful treatment of disease. An important area of research in disease detection lies in the use of magnetic resonance imaging (MRI), which provides three-dimensional images of deep tissues in a noninvasive manner. Molecular compounds that bind the lanthanide metal gadolinium (Gd³⁺) have traditionally been utilized as magnetic resonance imaging (MRI) contrast agents to increase the resolution of the tissue images because of the high relaxivity that these materials induce on water protons. We are interested in developing polymers and click clusters that have repeated Gd³⁺ chelates along the backbone. These materials may improve several properties of the molecular materials that are currently in use. Polymeric contrast agents may enhance MRI resolution, increase circulation lifetime, reduce toxicity, and enhance the tissue-specific concentration of Gd³⁺. Carbohydrate-based polymers that bind Gd³⁺ are currently being synthesized. We are interested in studying the structure-relaxivity relationships of different polymers and click cluster via inversion recovery experiments and evaluating toxicity profiles in vitro and in vivo. In addition, new methods are being developed to enhance and study tissue-specific uptake of these polymers.

Theranostic Agent Development

Smart biomaterials termed “theranostic” agents are being developed that provide diagnostic imaging, therapeutic delivery, and the ability to monitor treatment efficacy in real time. Indeed, the parallel development of novel nucleic acid drugs and theranostic vehicles that offer disease diagnosis, treatment, and the ability to understand the delivery mechanisms/kinetics on a range of biological scales will advance the field of nucleic acid therapeutics toward the discovery of personalized treatment strategies. To this end, lanthanide metals have been incorporated into our nucleic acid delivery vehicles. For example, europium's (Eu³⁺) red luminescence allows the detection of the delivery vehicle within cultured cells on the nanometer/micron scale. Likewise, gadolinium (Gd³⁺), which is routinely used as a MRI contrast agent is also being incorporated to monitor nucleic acid delivery in vivo on the tissue (sub-millimeter) scale.  Projects in this area are focused on the discovery of novel structures and understanding their efficacy/detection limits in biological systems.