• Develop new computational analyses to align different single cell datasets, including those generated by disparate laboratories.
• Develop new tools and methods to integrate and spatially align single cell data.
• Develop new ways of mining of clinical, histological, tissue, cellular and molecular data to create a 2- or 3-Dimensional tissue atlas or conceptual model that visualizes multi-level data and generates usable information.
• Develop new methods to register overlapping images of ultrastructure and molecular markers.
• Develop better ways to aggregate and integrate data and metadata from a broad range of experimental and computational projects.
• Develop and/or adapt data analysis tools for searching, integration, annotation, cross-validation and visualization.

• The transparency of C. elegans and zebrafish larvae permits the facile monitoring of cell-based biosensors designed to measure physiology and function in free living organisms. With the advent of improved genome-editing technologies and large-scale efforts to develop biosensors (e.g. ER stress, oxidative stress, hypoxia, autophagy, glucose levels, hormone levels, albuminuria, sheer stress, fluid flow, etc.), the time is right for researchers to develop novel tools to simultaneously image/measure physiologic processes in real-time.
• Develop a high-throughput method to measure transepithelial transport in vivo by adapting available ion, pH and/or volume sensitive fluorescent dyes. Such dyes could be engineered into cells or be applied to growing organoids.
• Develop assays of mucosal immune, gut barrier, and other intestinal functions which could be used to elucidate functional consequences of human mutations conferring diabetic complications risk.
• Develop reporter lines to track development of inflammation or fibrosis in liver, kidney, bladder and other end organs of diabetic complications.

• Develop non-viral methods to deliver CRISPR/Cas9 to specific cells for genome editing.
• Develop cell specific nanoparticles, extracellular vesicles, or exosomes to deliver nucleic acids (e.g., mRNA, siRNA), or develop tools to purify and characterize these delivery vehicles.
• Develop novel encapsulation technologies that enhance solubility and bioavailability, and facilitate cell-specific entry.

• Develop and apply a scalable technology for high spatial resolution -omic (e.g., transcriptomic, proteomic, metabolomic) interrogation of human tissue.
• Develop novel single cell proteomics methods.
• Use computational methods to extract multiple quantifiable image features to show differences in (patho)physiology, (dys)function, or response to intervention.
• Develop new analysis methods to interrogate biomedical image data.
• Develop imaging techniques to measure organ function and pathology (e.g., inflammation, altered metabolism, fibrosis) where other diagnostics are misleading/inadequate or to measure novel prognostic indicators that could be used to improve clinical trials.
• Produce, validate and distribute novel antibodies and probes based on single cell or other -omic datasets.
• Develop new ways of high-order mining of clinical, histological, tissue, cellular and molecular data to create a 2- or 3-Dimensional tissue atlas or conceptual model that visualizes multi-level data and generates usable information.
• Develop new methods to register overlapping images of ultrastructure and molecular markers.

• Develop (or adapt) experimental models of diabetic end-organ complications in both sexes.
• Leverage existing models for studying sex-differences to differentiate sex hormone from sex chromosome effects in diabetic end-organ complications.
• Perform exploratory studies to identify differentially expressed genes in diabetic males and females.
• Perform exploratory studies to identify sex-specific biomarkers for diabetic end-organ complications.

• Support the preclinical work necessary to obtain "proof of principle" establishing that a new molecule or novel approach will be a viable candidate for expanded clinical evaluation.
• Develop and use novel cell-based, organoid, or in vivo assays for the discovery and preclinical testing of potential therapies for diabetic complications.
• Perform additional preclinical testing as needed to validate potential therapies under disease-specific conditions and in multiple systems before they can progress along the drug development pipeline.
• Bioengineer healthy and diseased end organs of diabetic complications, including incorporation of appropriate cell types, for functional testing and screening.
• Identify and interrogate endogenous repair and regeneration pathways. Screen for ways to promote endogenous repair and regeneration in vivo.
• Define repair circuits to identify drug targets that will enhance productive repair and develop target-based small screening assays. Use phenotype screening assays to identify small molecules that will stimulate productive repair/regeneration and restore failing function.
• Develop cell-based therapies, including induced pluripotent stem cell (iPS) and regenerative therapies to repair and reverse complications. Develop methods for identifying, counting and eliminating potentially harmful components and cells from stem cell preparations.

• Demonstrate the relationship (if any) between the human or rodent microbiome and the initiation, severity, frequency, or resolution of complications-related phenotypes;
• Define the interplay between host and microbiome genetics and epigenetics in driving or modulating complications-related phenotypes;
• Explore how penetrance and/or severity of complications-related phenotypes may be affected by defined microbiota populations or their products;
• Examine whether naturally-occurring or synthetic phages may influence complications-related outcomes by altering resident microbiota populations.
• Proposed microbiome studies should incorporate a paragraph entitled "Rigor and Reproducibility" that outlines how the experimental design addresses and/or controls for key parameters that may affect rigor and reproducibility in microbiome research, such as: husbandry conditions, key biological variables (such as sex, circadian rhythms, body weight, physical activity) and diet.

• Develop non-viral methods to deliver CRISPR/Cas9 to specific cells for genome editing.
• Develop cell specific nanoparticles, extracellular vesicles, or exosomes to deliver nucleic acids (e.g., mRNA, siRNA).
• Develop novel encapsulation technologies that enhance solubility and bioavailability, and facilitate cell-specific entry.

• Develop and apply a scalable technology for high spatial resolution -omic (e.g., transcriptomic, proteomic, metabolomic) interrogation of human tissue.
• Develop novel single cell proteomics methods.
• Use computational methods to extract multiple quantifiable image features to show differences in (patho)physiology, (dys)function, or response to intervention.
• Develop new analysis methods to interrogate biomedical image data.
• Develop imaging techniques to measure organ function where other diagnostics are misleading/inadequate or to measure novel prognostic indicators that could be used to improve clinical trials.
• Produce, validate and distribute novel antibodies and probes based on single cell or other -omic datasets.
• Develop new ways of high-order mining of clinical, histological, tissue, cellular and molecular data to create a 2- or 3-Dimensional tissue atlas or conceptual model that visualizes multi-level data and generates usable information.
• Develop new methods to register overlapping images of ultrastructure and molecular markers.

• Develop (or adapt) experimental models of diabetic end-organ complications in females.
• Leverage existing models for studying sex-differences to differentiate sex hormone from sex chromosome effects in diabetic end-organ complications.
• Perform exploratory studies to identify differentially expressed genes in diabetic males and females.
• Perform exploratory studies to identify sex-specific biomarkers for diabetic end-organ complications.

• Understand changes in the parasympathetic and sympathetic tone in response to diabetes, and how this imbalance may be implicated in end-organ complications.
• Dissect contributions of the extrinsic autonomic dysfunction from intrinsic autonomic dysfunction (enteric nervous system, neuroendocrine cells) in gastrointestinal, bladder or urogenital diabetic complications.
• Determine contribution of renal innervation to initial hemodynamic responses of the kidney to hyperglycemia.
• Determine changes in renal, bladder, urogenital, or GI tract afferent signaling in response to diabetes, and how this may relate to various dysfunctions (i.e. urinary incontinence, changes in gut motility, hypertension, glomerular filtration rate changes).

• Support the preclinical work necessary to obtain "proof of principle" establishing that a new molecule or novel approach will be a viable candidate for expanded clinical evaluation.
• Pilot studies to support the development and use of novel cell-based, organoid, or in vivo assays for the discovery and preclinical testing of potential therapies for diabetic complications.
• Additional preclinical testing is often required to validate potential therapies under disease-specific conditions and in multiple systems before they can progress along the drug development pipeline.
• Bioengineer healthy and diseased end organs of diabetic complications, including incorporation of appropriate cell types, for functional testing and screening.
• Identify and interrogate endogenous repair and regeneration pathways. Screen for ways to promote endogenous repair and regeneration in vivo.
• Define repair circuits to identify drug targets that will enhance productive repair and develop target-based small screening assays. Use phenotype screening assays to identify small molecules that will stimulate productive repair/regeneration and restore failing function.
• Application of cells, including induced pluripotent stem (iPS) cells, and regenerative therapies to repair and reverse complications. Develop methods for identifying, counting and eliminating potentially harmful components and cells from stem cell preparations.

• Develop imaging techniques to measure organ function where other diagnostics are misleading/inadequate or to measure novel prognostic indicators that could be used to improve clinical trials.
• Develop imaging assays to detect tissue heterogeneity with prognostic significance.
• Develop new analysis methods to interrogate biomedical image data.
• Develop imaging techniques to resolve pathological targets and pathways that may be modulated with interventions (such as inflammation and fibrosis).
• Use computer extraction of multiple quantifiable image features to show differences in (patho)physiology, (dys)function, or response to intervention.
• Develop new imaging protocols, tools or reagents for interrogation of healthy and diseased human tissue with multiple probes ("multiplexing") and single cell resolution.
• Develop novel ways to non-invasively image productive repair and regeneration, perhaps by imaging the integration and/or "fusion" of applied stem cells.
• Produce, validate and distribute novel antibodies and probes based on single cell datasets.
• Spatial Integration and Visualization of Single Cell Data.

• Develop new computational analyses to align different single cell datasets.
• Develop new tools and methods to integrate and spatially align single cell data.
• Develop new ways of high-order mining of clinical, histological, tissue, cellular and molecular data to create a 2- or 3-Dimensional tissue atlas or conceptual model that visualizes multi-level data and generates usable information.
• Develop new methods to register overlapping images of ultrastructure and molecular markers.
• Develop better ways to aggregate and integrate data and metadata from a broad range of experimental and computational projects.
• Develop and/or adapt data analysis tools for searching, integration, cross-validation and visualization.
• Provide high-quality spatio-temporal annotation and develop ontologies. Extend existing ontologies to resolve interstitial components, cell type and cell state.

• Develop (or adapt) experimental models in which females develop diabetic end-organ complications. As females may be protected from diabetes due to estrogen-protective effect on pancreatic β-cells, females are often resistant to developing end-organ complications, which may explain why females are often overlooked in basic studies.
• Leverage existing models for studying sex-differences to differentiate sex hormone from sex chromosome effects in diabetic end-organ complications.
• Perform exploratory studies to identify differentially expressed genes in diabetic males and females.
• Perform exploratory studies to identify sex-specific biomarkers for diabetic end-organ complications.

• Understand changes in the parasympathetic and sympathetic tone in response to diabetes, and how this imbalance may be implicated in end-organ complications.
• Dissect contributions of the extrinsic autonomic dysfunction from intrinsic autonomic dysfunction (enteric nervous system, neuroendocrine cells) in gastrointestinal, bladder or urogenital diabetic complications.
• Determine contribution of renal innervation to initial hemodynamic responses of the kidney to hyperglycemia.
• Determine changes in renal, bladder, urogenital, or GI tract afferent signaling in response to diabetes, and how this may relate to various dysfunctions (i.e. urinary incontinence, changes in gut motility, hypertension, glomerular filtration rate changes).

• Support the preclinical work necessary to obtain "proof of principle" establishing that a new molecule or novel approach will be a viable candidate for expanded clinical evaluation.
• Pilot studies to support the development and use of novel cell-based, organoid, or in vivo assays for the discovery and preclinical testing of potential therapies for diabetic complications.
• Additional preclinical testing is often required to validate potential therapies under disease-specific conditions and in multiple systems before they can progress along the drug development pipeline.
• Bioengineer healthy and diseased end organs of diabetic complications, including incorporation of appropriate cell types, for functional testing and screening.
• Identify and interrogate endogenous repair and regeneration pathways. Screen for ways to promote endogenous repair and regeneration in vivo.
• Define repair circuits to identify drug targets that will enhance productive repair and develop target-based small screening assays. Use phenotype screening assays to identify small molecules that will stimulate productive repair/regeneration and restore failing function.
• Application of cells, including induced pluripotent stem (iPS) cells, and regenerative therapies to repair and reverse complications. Develop methods for identifying, counting and eliminating potentially harmful components and cells from stem cell preparations.

• Develop new protocols, tools or reagents for the molecular/omic interrogation of healthy or diseased human tissue. These may involve imaging methods at the single cell level (e.g., SWITCH, systems-wide control of interaction time and kinetics of chemicals, CyTOF mass cytometry, CLARITY, etc.) or digestion of specimens into single cells for further analysis (e.g, flow cytometry, microfluidics, mass cytometry or chemical cytometry).
• Profile and validate cell types isolated from healthy or diseased human tissue using FACS, laser-capture or other approaches. Molecular/omic profiling of cell types should inform in vitro and in vivo efforts to model, recreate or regenerate these cell types to study human (patho)physiology and (dys)function.
• Catalog cells and pathways involved in productive repair post diabetic injury.
• Assess the quality of human bone in diabetes. Increased fracture risk is an important morbidity in subjects with T2DM. Bone imaging technologies, have shown differences in bone quality and bone marrow fat content as compared to non-diabetic individuals. The role of advanced glycation end-product (AGE) accumulation has also been implicated in this increased fracture risk. This information has increased our understanding of how T2DM adversely impacts both bone metabolism and fracture risk, but the histological characterization of skeletal abnormalities in T2DM has been poorly explored. This information may provide important insights that may lead to a better understanding of this complication as well provide insights on how to ameliorate it.
• Use novel tissue interrogation techniques to better define the diabetic human prostate.
• Examine the localized tissue secretory and cellular microenvironments (e.g., extracellular matrix, interstitium, wound exudate) using emerging technologies such as nanoFACS for extracellular vesicle profiling, CLASI-FISH for exploring the 3D relationships in the microbiome, or MALDI-based technologies to characterize the extracellular proteome as it relates to the development or extent of diabetic complications.

• Develop tissue chip models of end organs affected by diabetic complications, including components that may be affected by diabetic complications (e.g., microvasculature or innervation).
• Use human cells to develop mature organoid models of end organs affected by diabetic complications. Develop methods to support the maturation of organoids.
• Study mechanisms of diabetic complications using a tissue chip model of an affected end organ.
• Evaluate a therapeutic strategy for diabetic complications using an assay involving an organoid model.
• Study impact of patient genotype on diabetic complications using tissue chip models incorporating human cells.

• Identify and interrogate endogenous repair and regeneration pathways. Screen for ways to promote endogenous repair and regeneration in vivo.
• Define repair circuits to identify drug targets that will enhance repair and develop target-based small screening assays. Use phenotype screening assays to identify small molecules that will stimulate repair/regeneration and restore failing function.
• Discover and/or characterize signals released by injured tissue required for productive repair and regeneration.
• Application of cells, including induced pluripotent stem (iPS) cells, and regenerative therapies to repair and reverse complications.
• Develop novel ways to non-invasively image repair and regeneration, perhaps by imaging the integration and/or "fusion" of applied stem cells.
• Develop methods for identifying, counting and eliminating potentially harmful components and cells from stem cell preparations.

• Develop non-mammalian model organism lines that mimic aspects of the human pathology through protocols that induce diabetes and/or express cell-based physiologically-relevant biosensors and reporters to simultaneously image/measure physiologic processes in real-time.
• Develop non-mammalian models organism lines and protocols to study the effects of altered glucose homeostasis on the central and peripheral nervous systems leading to further disruption of metabolic control and complications such as neuropathic pain, gastroparesis, and disturbed renal function.
• Develop non-mammalian model organism assays, reporters, or other tools to identify novel or understudied processes that impact the molecular anatomy and physiology of kidney and urinary tract development, function or dysfunction in response to long-term dysglycemia.
• Develop assays of mucosal immune, gut barrier, and other intestinal functions which could be used to elucidate functional consequences of human mutations conferring diabetic complications risk.
• Develop reporter lines to track development of fibrosis in liver, kidney and other end organs of diabetic complications.

• Support the preclinical work necessary to obtain "proof of principle" establishing that a new molecule or novel approach will be a viable candidate for expanded clinical evaluation.
• Pilot studies to support the development and use of novel cell-based, organoid, or in vivo assays for the discovery and preclinical testing of potential therapies for diabetic complications.
• Additional preclinical testing is often required to validate potential therapies under disease-specific conditions and in multiple systems before they can progress along the drug development pipeline.
• Bioengineer healthy and diseased end organs of diabetic complications, including incorporation of appropriate cell types, for functional testing and screening.

• Develop new protocols, tools or reagents for the molecular/omic interrogation of healthy or diseased human tissue. These may involve imaging methods at the single cell level (e.g., SWITCH, systems-wide control of interaction time and kinetics of chemicals, CyTOF mass cytometry, CLARITY, etc.) or digestion of specimens into single cells for further analysis (e.g, flow cytometry, microfluidics, mass cytometry or chemical cytometry).
• Profile and validate cell types isolated from healthy or diseased human tissue using FACS, laser-capture or other approaches. Molecular/omic profiling of cell types should inform in vitro and in vivo efforts to model, recreate or regenerate these cell types.
• Examine the localized tissue secretory and cellular microenvironments (e.g., extracellular matrix, interstitium, wound exudate) using emerging technologies such as nanoFACS for extracellular vesicle profiling, CLASI-FISH for exploring the 3D relationships in the microbiome, or MALDI-based technologies to characterize the extracellular proteome as it relates to the development or extent of diabetic complications.
• Bioengineer healthy or diseased end organs of diabetic complications, including incorporation of appropriate cell types, for functional testing and screening.

• Develop non-mammalian model organism lines that mimic aspects of the human pathology through protocols that induce diabetes and/or express cell-based physiologically-relevant biosensors and reporters to simultaneously image/measure physiologic processes in real-time.
• Develop non-mammalian models organism lines and protocols to study the effects of altered glucose homeostasis on the central and peripheral nervous systems leading to further disruption of metabolic control and complications such as neuropathic pain, gastroparesis, and disturbed renal or cardiac function.
• Develop non-mammalian model organism assays, reporters, or other tools to identify novel or understudied processes that impact the molecular anatomy and physiology of kidney and urinary tract development, function or dysfunction in response to long-term dysglycemia.
• Develop assays of mucosal immune, gut barrier, and other intestinal functions which could be used to elucidate functional consequences of human mutations conferring diabetic complications risk.
• Develop reporter lines to track development of fibrosis in liver, kidney and other end organs of diabetic complications.

• Biofilms can be monolayer or multilayer and contain heterogeneous or homogenous populations of microorganisms associated with biotic and abiotic surfaces. Biofilm formation by pathogenic bacteria contribute significantly to antibiotic resistance and infection recurrence. Host-pathogen interaction, extracellular matrix formation, and quorum sensing are involved in the formation of biofilms but little is known about how these processes are affected by changes in tissue and cellular physiology in the context of chronic diabetes. Such changes may contribute to diabetic complications such as recurrent urinary tract infections, bladder dysfunction and prostatitis.
• It has become increasingly apparent that the urine is not a "sterile" environment. Recent data indicate the bladder and urine are hosts to a rich array of micro-organisms that are likely to play important roles in maintaining normal bladder function. We encourage pilot proposals exploring how these "native" micro-organisms interact with the pathogenic micro-organisms to promote or prevent biofilm formation and recurrent UTIs within the context of diabetes.
• Artificial surfaces commonly used for in-dwelling urinary catheters are a recognized substrate for biofilm formation and the unique milieu of diabetes may alter the population of micro-organisms present in the urinary bladder to promote biofilm formation. Specific topics of interest include elucidating the interaction of pathogenic micro-organisms with catheter materials and the exploration of alternative materials that preclude pathogenic biofilm formation in diabetic patients.
• The twenty-week healing rate for neuropathic diabetic foot ulcers is about 50% and bacterial biofilms are a major cause for the failure of chronic wounds to heal. Detection of biofilms is critical for diagnosis, but standard culture methods and other indicators such as visual and olfactory examination of wounds are inadequate. The treatment of diabetic foot ulcers requires a better understanding of the mechanisms by which certain bacteria within the complex flora of a diabetic wound and within biofilms become pathogenic. New molecular microbiological technologies have the potential to detect bacterial biofilms and their role in the chronicity of diabetic foot ulcers. Specific topics of interest are the discovery and study of novel pathogenic mechanisms of biofilms for diabetic wound healing and the development of detection methods for biofilms in diabetic wounds for use in research and the clinic.

Insulin resistance and type 2 diabetes impacts neurocognitive function via mechanisms independent from those typically associated with Alzheimer's Disease pathology. In addition, there is a growing appreciation that type 1 diabetes may be associated with neurocognitive dysfunction. Understanding the mechanism(s) that explain the neurocognitive complications of insulin resistance and diabetes (both type 1 and type 2) will be important if we are going to develop novel therapeutic targets and approaches. Moreover, an acute clinically important feed-forward downward spiral can develop because even mild neurocognitive impairment can prevent impaired persons with diabetes from managing his/her anti-diabetic medications. Thus, there are important individual, societal, and economic implications.
Metabolic disease may impact the brain in ways that differ from its effects on other end organs of diabetic complications. It is possible that due to aging and disease-related factors, isolating mechanisms may be more complex and clinical targets less modifiable as individuals advance in age and disease burden increases. Therefore, there will be a special emphasis on research that addresses potential mechanisms that may be independent of those associated with aging or complex disease.

• Development, translation, and/or validation of imaging or other methods to measure the neuropathophysiologic changes associated with diabetes.
• Elucidating mechanisms for how diabetes may impact the brain in ways that differ from its effects on other end organs.
• Effect of neuropathology and brain dysfunction on peripheral metabolism and diabetes.
• Clinicopathological correlation as well as cause-and-effect research designs aimed at identifying the sequence(s) of events that can lead to the development of specific neural systems and cognitive functions impacted by diabetes and its other complications, especially disruptions with the great impact on clinical care.
• Approaches that take advantage of recent breakthroughs in the genetics of diabetes and insulin resistance are encouraged, as are computational approaches applied to multi-omics brain data from persons with diabetes.

• Support the preclinical work necessary to obtain "proof of principle" establishing that a new molecule or novel approach will be a viable candidate for expanded clinical evaluation.
• Pilot studies to support the development and use of novel cell-based, organoid, or in vivo assays for the discovery and preclinical testing of potential therapies for diabetic complications.
• Additional preclinical testing is often required to validate potential therapies under disease-specific conditions and in multiple systems before they can progress along the drug development pipeline.
• Bioengineer healthy and diseased end organs of diabetic complications, including incorporation of appropriate cell types, for functional testing and screening.

• Develop non-mammalian model organism lines that a) mimic aspects of the human pathology of diabetic complications through protocols that induce diabetes and b) express cell-based physiologically-relevant biosensors and reporters to simultaneously image/measure physiologic processes in real-time to explore the sequence of inter- and intra-cellular events leading to cellular dysfunction, disturbed physiological processes, and end-organ damage.
• Develop non-mammalian models organism lines and protocols to study the effects of altered glucose homeostasis on the central and peripheral nervous systems leading to further disruption of metabolic control and complications such as neuropathic pain, gastroparesis, and disturbed cardiac function.
• Develop non-mammalian model organism assays, reporters, or other tools to identify novel or understudied processes that impact the molecular anatomy and physiology of kidney and urinary tract development, function or dysfunction in response to long term dysglycemia.
• Develop assays of mucosal immune, gut barrier, and other intestinal functions which could be used to elucidate functional consequences of human mutations conferring diabetic complications risk.
• Develop reporter lines to track development of fibrosis in liver, kidney, adipose tissue and other end organs of diabetic complications.
  
  

• Sustaining a healthy population of mitochondria requires the addition of new functional mitochondria and removal of old dysfunctional mitochondria. In addition to de novo biogenesis, circulating stem cells have been shown to share their functional mitochondria with injured cells (including hepatocytes, lung epithelia, and hematopoietic stem cells) to mitigate tissue damage. Conversely, recent evidence suggests that retinal neurons can package their dysfunctional mitochondria and expel them to neighboring astrocytes for degradation (transmitophagy).
• Mitochondrial bioenergetics has been implicated in a broad spectrum of metabolic and degenerative diseases. A new class of drugs targeting cardiolipin is believed to improve mitochondrial "plasticity" and is being tested in diabetes, heart failure, ischemic injury, neurodegeneration, retinal and skin diseases, and chronic kidney disease.
• Adult stem cells remain metabolically inactive until triggered to proliferate and replenish a tissue. Recent work suggests that the mitochondrial unfolded protein response is coupled to cellular metabolism and regulates stem cell aging. In hematopoietic stem cells, for example, it appears that aging is not due to passive accumulation of cellular damage over time, but rather an active repression of protective pathways and suggests that targeting these protective pathways in mitochondria could reverse aging and restore tissue homeostasis.

• Diabetic foot ulcers are a common complication of diabetes with a lifetime prevalence of about 25%. The twenty week healing rate for neuropathic diabetic foot ulcers is about 50% and bacterial biofilms are a major cause for the failure of chronic wounds to heal. Detection of biofilms is critical for diagnosis, but standard culture methods and other indicators such as visual and olfactory examination of wounds are inadequate. The treatment of diabetic foot ulcers requires a better understanding of the mechanisms by which certain bacteria within the complex flora of a diabetic wound and within biofilms become pathogenic. New molecular microbiological technologies have the potential to detect bacterial biofilms and their role in the chronicity of diabetic foot ulcers. Specific topics of interest are the discovery and study of novel pathogenic mechanisms of biofilms for diabetic wound healing and the development of detection methods for biofilms in diabetic wounds for use in research and the clinic.
• Biofilms can be monolayer or multilayer and contain heterogeneous or homogenous populations of microorganisms associated with biotic and abiotic surfaces. Biofilm formation by pathogenic bacteria contribute significantly to antibiotic resistance and infection recurrence. Host-pathogen interaction, extracellular matrix formation quorum sensing are involved in the formation of biofilms but little is known about how these processes are affected by changes in tissue and cellular physiology due to diabetes. Such changes may contribute to diabetic complications such as recurrent urinary tract infections and bladder dysfunction.

• A recent meta-analysis of T1DM observational studies suggests that, compared to nondiabetic individuals, women with Type 1 diabetes have twice the excess risk of fatal and non-fatal cardiovascular events than men with Type 1. Similar evidence supports the same in women with Type 2. The biological mechanisms underlying this difference are unclear.
• Men with increased adiposity and Type 2 diabetes have lower testosterone levels and higher circulating estrogens due to somatic aromatase activity. These levels of estrogen can predispose men to urologic complications. Few studies have examined these differences mechanistically, but estrogen signaling through both genomic and non-genomic receptors appears to play a role.

• The shared pathophysiology of diabetes and dementia, including vascular and Alzheimer's disease pathology (e.g. Aβ, tau).
• Pathophysiology unique to diabetes (e.g. glycemia, insulin) and the impact on cognition and dementia.
• The effect of neuropathology and brain dysfunction on peripheral metabolism and diabetes.

• Support the preclinical work necessary to obtain "proof of principle" establishing that a new molecule or novel approach will be a viable candidate for expanded clinical evaluation.
• Pilot studies to support the development and use of novel cell-based, organoid, or in vivo assays for the discovery and preclinical testing of potential therapies for diabetic complications.
• Additional preclinical testing is often required to validate potential therapies under disease-specific conditions and in multiple systems before they can progress along the drug development pipeline.