Using a powerful analytical technique known as ChemoMorphic Analysis, INTiDYN evaluates small biopsies from animals or humans to create a morphological, biochemical, and genomic snapshot of gender, age, disease, drug, or toxin impacts in complex tissue. Interpretation of these analyses yields critical information about tissue transformations and molecular/cellular interactions, as well as the effects on the various functional systems integrated within the skin (e.g., cardiovascular, nervous, endocrine, immunological).

CMA

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a
Clinical Assessment and Skin Biopsy

Clinical sensory testing is performed, followed by the collection of 3mm skin biopsies which are immediately immersion fixed or flash frozen.
b
Biopsy Sectioning and Sequential Distribution

Immersion fixed and flash frozen biopsies are cut into sections and sequentially mounted onto a series of slides for immunochemistry or laser capture. This method of tissue distribution ensures that each slide contains sections throughout the biopsy. A mix of section thicknesses and number of sections per slide optimizes tissue use and maximizes options for biomarker combinations.
c
Multimolecular Immunolabeling

Individual slides are processed for immunofluorescent detection of protein expression using a variety of antibody combinations – those desired by the client and/or recommended by INTiDYN .
d
Compartmentalized Qualitative and Quantitative Microscopic Analysis

Immunolabeled slides are evaluated under epifluorescence wide-field and confocal microscopy. Individual tissue compartments undergo qualitative assessments and quantitative measures of endpoints chosen by our clients and/or recommended by INTiDYN .
e
Compartmentalized Laser Capture Microdissection

Flash frozen sections are collected on slides for laser capture microdissection (LCM) of individual skin compartments, such as epidermis. Compartment specific etchings from sections throughout the biopsy are pooled for nucleic acid extraction.
f
Nucleic Acid Extraction and Functional Genomic Assays

Total genomic DN A (gDN A), micro RNA (miRNA), and messenger RNA (mRNA) are recovered for array analysis or quantitative PCR (qPCR) to the specifications of our clients and/or at the recommendation of INTiDYN .
g
Data Compilation

INTiDYN scientists utilize all levels of data generated within the scope of a project (clinical, cellular/molecular, genetic) to determine:

Potential disease mechanisms and therapeutic targets

Efficacy and/or off-target toxicity of potential therapeutics

Impacts of normal demographic variables such as gender, race, and age on disease mechanisms and potential therapeutics

Integrated Tissue Dynamics (INTIDYN) has applied their capabilities to create solutions for many types of customers spanning across Academic Institutions, Pharmaceutical and Biotechnology Companies and other Medical Professions. Below are three case studies highlighting some of the ways in which INTIDYN has applied their analysis and advisory services.

Case Study 1: Preclinical Detection of a Severe Side Effect
Background
  • Nerve injuries have been shown to commonly result in severe pain caused by stimuli that are not normally painful (i.e. allodynia).
  • Published studies have shown that this condition is accompanied by alterations in epidermal sensory innervation that expresses the neuropeptides CGRP and Substance P.
INTIDYN's Assignment
  • Our client developed a novel therapeutic that they hypothesized would prevent or reverse this pathology. INTIDYN was asked to determine if this hypothesis was accurate.
Result
  • ITD-CMA confirmed that the therapeutic was successfully impacting the predicted innervation.
Value-Added Interpretative Result Based on ITD-CMA:
  • However, we first unblinded the study by detecting that the therapeutic was profoundly disrupting the structural integrity of the cutaneous arterioles.
  • In this case we were able to warn our client that their promising therapeutic had a potentially serious vascular side-effect.

 

Case Study 2: Rectifying an Incorrect Hypothesis Redirects a Therapeutic Strategy
Background
  • Several studies have documented that pain and numbness caused by chemotherapy is accompanied by a loss of cutaneous innervation.
  • Our client developed a novel therapeutic that was designed to prevent the loss of cutaneous innervation caused by a standard chemotherapy regimen.
INTIDYN's Assignment
  • Determine if our client’s therapeutic agent was limiting chemotherapeutic damage to cutaneous innervation.
Result
  • Using different immunochemical markers from those routinely used to detect sensory endings, our ITD-CMA revealed that the chemotherapy was not causing a loss of innervation but was, in fact, causing a down regulation among the standard neuronal biomarkers.
Value-Added Interpretative Result Based on ITD-CMA:
  • This finding shifted the course of the study from attempts to prevent neuronal loss to strategies that would restore normal neuronal biochemistry.

 

Case Study 3: Discovering Previously Unknown Therapeutic Targets
Background
  • Many studies of humans with various types of painful peripheral neuropathies have documented a loss of intraepidermal nerve fibers.
  • This has led to a hypothesis that the chronic pain may be due to an increase of spontaneous activity among neurons in the spinal cord due to a loss of sensory input (i.e. deafferentation syndrome).
INTIDYN's Assignment
  • Our client was running a human study that combined physiological assessments of sensory nerve function (microneurography) with our ITD-CMA of cutaneous innervation to assess a wide variety of neuropathic conditions.
Result
  • The purpose of the study was to lay the groundwork for future therapeutic strategies to treat neuropathic pain by promoting regeneration of cutaneous innervation.
Value-Added Interpretative Result Based on ITD-CMA:
  • However, our results also revealed that a comparable loss of intraepidermal nerve fibers also occurred in neuropathies that were not painful.
  • These results demonstrated that a loss of epidermal nerve fibers was not necessarily the cause of chronic pain.
  • In the course of the study we did discover previously unknown changes in other biomarkers that more consistently correlated with the presence of neuropathic pain.

INTIDYN offers expert analysis and interpretation.

Discover Development Translation

 

  • Traditional approaches have produced few effective therapeutics for neuropathies and dermatopathologies. These are typically transient and prone to severe side effects.
  • In the absence of tissue structure, the power of traditional approaches is limited.
  • By analyzing tissue structure and chemistry, Intidyn is able to address those questions that were ambiguous under traditional approaches.

Intidyn's core competency is the design, execution, and interpretation of immunochemical structural analyses of tissue specimens, particularly skin biopsies spanning across both basic and applied research settings. Intidyn has extensive experience with preclinical and clinical studies. Confidentiality will be maintained according to the clients' needs and industry standards.

Study design: Intidyn will assist the client in designing research projects that the client wishes to conduct with their own personnel or that they wish to have Intidyn conduct.

Execution: A typical project will involve sectioning, immunochemical processing, and microscopic analysis of skin specimens according to the needs of the client. The specimens can be provided by the client and shipped to Intidyn according to the specifications of Intidyn. Intidyn can also recommend or arrange collaborations for potential alternate sources of specimens, such as qualified clinicians for clinical studies. Immunochemical assessments will be conducted by epifluorescence and/or confocal microscopy using systems and software engineered by MBF Biosciences. Assessments can be qualitative and quantitative according to our clients needs. Publication quality reports on the results will be provided.

Interpretation: Intidyn can assist the client in interpreting the results of studies conducted by the client or by Intidyn on behalf of the client. The interpretation will be based upon Intidyn's vast experience in integrating and interpreting results from numerous previously published studies and unpublished Intidyn proprietary information.

Additional services: Intidyn can arrange to share portions of skin specimens in collaboration with other investigators who have expertise in other technologies such as assessments of message levels using RTPCR, in situ hybridization and gene arrays.

For more information on how INTIDYN has applied their capabilities to create solutions for their customers please click on the Case Studies link in the left menu bar.