Four in one—Combination therapy using live Lactococcus lactis expressing three therapeutic proteins for the treatment of chronic non-healing wounds

Abstract

Diabetes mellitus is one of the major concerns for health care systems, affecting 382 million people worldwide. Among the different complications of diabetes, lower limbs chronic ulceration is a common, severe and costly cause of morbidity. Diabetic foot ulcers are a leading cause of hospitalization in diabetic patients and its rate exceed the ones of congestive heart failure, depression or renal disease. Diabetic non-healing ulcers account for more than 60% of all non-traumatic lower limb amputations and the five-year mortality after amputation is higher than 50%, being equal to several types of advanced cancer. The primary management goals for an existing diabetic foot ulcer are to achieve primary healing as expeditiously as possible and to achieve a reduction of the amputation rate in the patients. Unfortunately, approximately a quarter of patients do not partially or fully respond to the standard of care. Advanced therapies for chronic wounds are existing, however, recent guidelines including the latest reviews and meta-analyses of the scientific and clinical evidence available from current treatment strategies and new therapeutic agents revealed that there is a lack of clinical data and persistent gap of evidence for many of the advanced therapeutic approaches. In addition, no pharmacological wound healing product has gained authority approval for more than 10 years in both US and EU, constituting a highly unmet medical need. In this publication we present data from a live biopharmaceutical product AUP1602-C designed as a single pharmaceutical entity based on the non-pathogenic, food-grade lactic acid bacterium Lactococcus lactis subsp. cremoris that has been genetically engineered to produce human fibroblast growth factor 2,interleukin4 and colony stimulating factor 1. Designed to address different aspects of wound healing (i.e. fibroblast proliferation, angiogenesis and immune cell activation) and currently in phase I clinical study, we show how the combination of the individual components on the wound micro-environment initiates and improves the wound healing in chronic wounds.

Engineered bacteria to accelerate wound healing: an adaptive, randomised, double-blind, placebo-controlled, first-in-human phase 1 trial

“… 20 So far, two attempts have been reported to accelerate healing in mouse models: Lactococcus lactis expressing FGF2, CSF1, and IL-4 (AUP-1602-C) currently tested in a first-human trial ( NCT04281992 ), and the herein investigated ILP100-Topical, L. reuteri R2LC expressing CXCL12. 11 , 21 In addition to the onsite bacterial production, the lactic acid produced by L. reuteri R2LC was demonstrated to reduce CXCL12 degradation within the wound, and thereby further boosting the CXCL12-induced tissue restorative functions of macrophages. 11 Accelerated healing of wounds by ILP100-Topical was also confirmed in minipigs.…”

Live Biotherapeutics : importance of formulation and lyophilization parametersand an example of a clinical application

“…Tolerability, clinical and biologic effects were assessed at each visit (SAD: Day 1, 2, 3, 7, 14, and at 6 weeks, 3 months and 12 months from start of treatment at Day 1; MAD: Day 1,2,3,5,8,10,12,15,17,19,21,32, and at 6 weeks, 3 months and 12 months after last dose at Day 19). All assessments were blinded and occurred by on-site visual inspections of the wounds by the Principal Investigator (or co-investigator), as well as off-site by traceable evaluation and detailed wound area measurements from 2D photographs of all wounds by 3 Independent Evaluators (IEs) with expertise in wound healing.…”

Signaling pathways in cutaneous wound healing

“…The last factors influencing wound healing are GSF (granulocyte stimulating factor) and VEGF (vascular endothelial growth factor). The latter was significantly reduced during wound healing (by 50%) in an experiment performed on diabetic rats compared to healthy controls (Kurkipuro et al, 2022).…”