Structural analysis of a micron-sized deposit of Cu in an insulin ball from a person with diabetes.

BACKGROUND

Protein misfolding is a hallmark of aging, resulting in diabetes and neuroendocrine disorders. Insulin preparations also form aggregates known as insulin balls. Aggregated insulin preparations usually form amyloids and are stable in subcutaneous tissue, some specimens are cytotoxic to cultured cells.

METHODS

A multilayered structural analysis of the detailed morphology of 21 insulin balls was performed by connecting regions of interest along spatial axes. Gross and optical microscopic findings, Raman spectrometric analysis using formalin-fixed paraffin-embedded block specimens, matrix-assisted laser desorption/ionization-time of flight-mass spectrometry, microfocus X-ray computed tomography, scanning electron microscopy-energy dispersive X-ray spectroscopy analysis, and transmission electron microscopy analysis were performed.

RESULTS

Here, we show the presence of 100 µm Cu within an insulin ball removed from a woman in her mid-40s with diabetes. The insulin ball is made of insulin lispro in an insoluble state in the lower abdominal subcutaneous adipose tissue. Transmission electron microscopy reveals fibrous structures. Microfocus X-ray computed tomography detects many spots with strong light contrast in the insulin ball. Scanning electron microscopy-energy dispersive X-ray spectroscopic analysis shows that the largest light spot is elemental metallic copper without an oxidation state (Cu).

CONCLUSIONS

The largest amount of Cu found in living things is in a human. Our discovery of 100 µm Cu within the insulin ball supports the idea that insulin preparations from outside can disrupt the balance of metals, including Cu. In 2025, the patient continues to inject subcutaneous insulin preparations, but no new insulin balls appear.