HR001122S0022.pdf

Broad Agency Announcement Cornerstone

BIOLOGICAL TECHNOLOGIES OFFICE

HR001122S0022

June 2, 2022

HR001122S0022, Cornerstone

TABLE OF CONTENTS

PART I: OVERVIEW INFORMATION

PART II: FULL TEXT OF ANNOUNCEMENT

1. Funding Opportunity Description

1.1. Program Overview

1.2. Technical Areas

1.3. Technical Approach

1.4. Program Schedule, Milestones & Metrics

1.5. General Requirements

2. Award Information

2.1. General Award Information

2.2. Fundamental Research

3. Eligibility Information

3.1. Eligible Applicants

3.2. Organizational Conflicts of Interest

3.3. Cost Sharing/Matching

4. Application and Submission Information

4.1. Address to Request Application Package

4.2. Content and Form of Application Submission

4.3. Funding Restrictions

4.4. Other Submission Information

5. Application Review Information

5.1. Evaluation Criteria

5.2. Review of Proposals

6. Award Administration Information

6.1. Submission Status Notifications

6.2. Administrative and National Policy Requirements

6.3. Reporting

6.4. Electronic Systems

7. Agency Contacts

8. Other Information

8.1. Proposers Day

9. APPENDIX 1 – Volume II checklist

10. APPENDIX 2 – Addendum Request Form

PART I: OVERVIEW INFORMATION

Federal Agency Name – Defense Advanced Research Projects Agency (DARPA), Biological Technologies Office (BTO)

Funding Opportunity Title – Cornerstone Announcement Type – Initial Announcement Funding Opportunity Number – HR001122S0022 North American Industry Classification System (NAICS) – 541714 Catalog of Federal Domestic Assistance Numbers (CFDA) – 12.910 Research and

Technology Development Dates o Posting Date: June 2, 2022 o Proposers Day: June 7, 2022 https://sam.gov/opp/5a0592fa6bf941659a0b7c2591b6e2b4/view o Proposal Abstract Due Date and Time: June 30, 2022, 4:00 PM ET o Full Proposal Due Date and Time: August 18, 2022, 4:00 PM ET o BAA Closing Date: August 18, 2022

Concise description of the funding opportunity: The Cornerstone program will develop safe and effective Integrated Countermeasures (i.e., countermeasure and delivery system) to prevent brain injury. Military personnel face a high risk of traumatic brain injury (TBI), resulting in a debilitating long-term burden both to the person as well as the healthcare infrastructure. At this time, no treatments for TBI have progressed past Phase II clinical trials. Cornerstone will improve mission execution and warfighter protection by addressing this gap. Cornerstone will consist of two technical areas: (TA1) Identify, optimize, and validate prophylactic countermeasures that prevent relevant targets at sites of injury from initiating injury response(s) to kinetic injury (e.g., blast); and (TA2) Develop clinically relevant spatial and temporal delivery of countermeasures. Ultimately, the development of prophylactic therapeutics that prevent injury by blocking the initiation of harmful signaling [initiating at the sites of injury] will sustain warfighter readiness and facilitate a strategic, operational, and tactical advantage.

Anticipated individual awards – Multiple awards are anticipated.

Types of instruments that may be awarded – Procurement contract or other transaction agreements for prototypes.

Agency contact

The BAA Coordinator for this effort may be reached at:

Cornerstone@darpa.mil

DARPA/BTO

ATTN: HR001122S0022

675 North Randolph Street Arlington, VA 22203-2114 https://sam.gov/opp/5a0592fa6bf941659a0b7c2591b6e2b4/view mailto:Cornerstone@darpa.mil

PART II: FULL TEXT OF ANNOUNCEMENT

1. Funding Opportunity Description

This publication constitutes a Broad Agency Announcement (BAA) as contemplated in Federal Acquisition Regulation (FAR) 6.102(d)(2) and 35.016 and 2 C.F.R. § 200.203. Any resultant award negotiations will follow all pertinent law and regulation, and any negotiations and/or awards for procurement contracts will use procedures under FAR 15.4, Contract Pricing, as specified in the BAA.

The Defense Advanced Research Projects Agency (DARPA) is soliciting innovative proposals to address the development of safe and effective Integrated Countermeasures (i.e., delivery system and countermeasure) to prevent brain injury. Research will be focused on the following areas: (1) Identification, optimization, and validation of prophylactic countermeasures that prevent energetically responsive receptors (i.e., targets) from initiating injury response(s) to kinetic insult(s) inclusive of blast or impact; and (2) Development of Integrated Countermeasures with precise delivery (i.e., spatial and temporal control). Proposed research should investigate approaches that enable revolutionary advances in the prevention of traumatic brain injury (TBI).

Proposed approaches should investigate the prevention of the initiation of harmful signaling cascades at sites of injury, for example, in the central and/or peripheral nervous systems (CNS, PNS) and/or vasculature, and develop countermeasures that can be administered prophylactically (i.e., before exposure) to prevent behavioral or cognitive deficit from TBI. By program completion, Cornerstone’s Integrated Countermeasures will be capable of instantaneous (seconds post-exposure) protection for the operator against Department of Defense (DoD)-relevant kinetic injury, both from a molecular cascade standpoint, and cognitive and behavioral effect.

Specifically excluded is research that involves:

1. Genetic manipulation of the host in order to aid target engagement or achieve altered downstream signaling as a countermeasure.

2. Sole use of in vitro or in vivo model systems that are not accepted as representative of kinetic (e.g., blast or impact) injury modes.

3. Treatment strategies focused on interventions delivered beyond five seconds post-injury, with preference for pretreatment-based countermeasures that provide a prophylactic effect.

4. Reliance on invasive medical procedures for Integrated Countermeasure delivery (i.e., pumps, implants, lumbar or cranial injections).

5. Remotely triggered Integrated Countermeasure delivery (i.e., strategies that require user or medic administration post-injury).

6. Approaches not inclusive of real-time and/or highly resolved temporal and spatial techniques to validate targeting, safety, and efficacy of countermeasures.

Proposals that employ the approaches described in the above list may be deemed non-conforming and may not be considered for review.

1.1. PROGRAM OVERVIEW

The goal of the Cornerstone program is to develop an Integrated Countermeasure (i.e., countermeasure and delivery system) that, when administered prophylactically, protects against behavioral and cognitive injury in the context of blast- or impact-induced TBI. Integrated Countermeasures would do this by preventing the initiation of harmful signaling cascades through spatially and temporally relevant countermeasure delivery.

There is no Food and Drug Administration (FDA) approved, pharmacological treatment for TBI, leaving rehabilitation protocols as the only option for individuals diagnosed with TBI. Current research in TBI countermeasure development focuses on pharmacological approaches that target a single, late-stage (hours- to days- post-exposure) cellular or molecular event in the background of a multimodal, complex injury response that initiates within seconds post-exposure. This puts any post-exposure treatment in the position of needing to treat multiple injury processes simultaneously to achieve efficacy. Balancing the ability to engage many targets while minimizing off-target effects has presented an insurmountable challenge to all therapies designed for TBI post-exposure treatment. Additionally, even potentially effective and safe treatments may need to cross the blood brain barrier to engage the central nervous system (CNS), which has been an obstacle for many drug delivery and neuropharmacological interventions.

While pharmaceutical-based treatment options are nonexistent for TBI, prevention of TBI is also reliant upon antiquated technology. Currently, the only options for protection against TBI are physical protection measures, such as armored vehicles and bulky helmets, which fail to provide complete protection from injury.

To shift the paradigm, proposals are required to focus on the immediate (i.e., within the first five seconds) biological responder(s) to kinetic insult. Within this time frame there are limited molecular structures with the ability to directly respond to energetics. Energetically responsive receptors include mechano-, voltage-, or sensory-sensitive and are located in the CNS, peripheral nervous system (PNS), and vasculature. These energetic responsive receptors (e.g., targets), therefore, may not only sense the first biological responder(s) in the milliseconds to seconds after kinetic injury but then initiate the complex injury cascade that results in cognitive and physical deficits.

To address the shortcomings of current research (which is unable to adequately characterize these initial responses to drive countermeasure development), proposers are encouraged to embrace emerging technologies in neuroscience that have enabled real-time measurement and visualization of molecular and cellular responses in vitro, ex vivo, and in vivo. For example, technologies that enable the observation of energetically responsive receptors in real time would be extremely valuable in validating countermeasure strategies. Cornerstone performers will successfully pair these highly time-resolved observation capabilities with kinetic exposure models to evaluate causal linkages between interventions and outcomes post-exposure (as defined by validated assays), as part of a comprehensive countermeasure validation strategy.

Cornerstone technologies will address two technical areas (TAs), ultimately developing preventative countermeasures to TBI:

TA1: Identify and validate countermeasures that prevent relevant targets from initiating downstream signaling injury response(s) to kinetic injury.

TA2: Develop clinically relevant countermeasure delivery systems that prevent molecular targets from responding to kinetic insult at sites of injury (e.g., CNS, PNS, vasculature, and organ systems).

At program completion, Cornerstone will have developed Integrated Countermeasures (i.e., countermeasure and delivery system) that, at minimum, allow for repeated prophylactic administration, a three-day effective residence time per dose, and provide protection in response to injury at any time within the three-day window. Upon kinetic injury (e.g., blast), Integrated Countermeasures should demonstrate spatial and temporal bioactivity necessary to inhibit the initiation of injury processes (e.g., molecular pathway activation) and protect against the development of canonical neuropathological deficits associated with TBI.

Approaches should be tested iteratively and optimized against various challenges with increasing complexity and performance requirements as the program progresses (see Tables 1 and 2).

Proposals to the Cornerstone program should include approaches to achieve the following:

1. Structure-guided design utilizing high-resolution models of targets (e.g., mechanical, electrical, chemical responsive receptors and channel subtype(s)).

2. Countermeasures with inhibitory signaling downstream of the primary target.

3. Demonstration of strong and specific binding of countermeasures with selected target in vitro.

4. Demonstration of spatially and temporally controlled delivery (i.e., injury triggered delivery if appropriate, contextual specificity of temporal and spatial resolution).

5. Demonstration of Integrated Countermeasures without behavioral or physiological anomalies at relevant, repeated, prophylactic doses in the absence of kinetic exposure.

6. Demonstration of Integrated Countermeasures with clear protective effects using robust and widely accepted cognitive and behavioral animal model systems.

Performer test protocols (safety, efficacy) will be required to place Cornerstone-developed approaches on a path towards clinical translation and regulatory approval. Performers will be expected to submit regulatory applications to the FDA by the end of the program. Further, performers will be required to engage with the FDA and other U.S. Government regulatory groups for design guidance within Phase I of the program.

1.2. TECHNICAL AREAS

The Cornerstone program is structured as a five (5) year effort consisting of three (3) phases:

(Phase I (Base – 24 months), Phase II (Option One – 24 months), and Phase III (Option Two – 12 months)).

DARPA will utilize an Independent Verification and Validation (IV&V) testing infrastructure to validate and assess protection efficacy of performer-generated technologies.

Proposed efforts must describe a plan to address both of the TAs over the entire program duration (60 months). At the end of Phase II (48 months), performers will have developed a safe and effective Integrated Countermeasure capable of providing > 95% protection against blast-induced molecular, cognitive, and behavioral injury in a minimum of two genetically distinct in vivo models. See Program Schedule, Metrics and Milestones for additional details.

Tables 1 and 2 describe the schedule of milestones and program evaluations that will drive programmatic decisions.

The technical areas are described below:

1.2.1. TA1: Identify, Optimize, and Validate Prophylactic Countermeasures

Performers will identify, optimize, and validate approaches that counter the initiation of injury signaling cascades at sites of injury (e.g., CNS, PNS, vasculature) protecting against TBI.

Approaches to countering the initiation of pathophysiological pathways may include novel small molecules, repurposed pharmaceuticals, natural products, immunotherapies, cell therapies, and/or other approaches capable of preventing targets from initiating molecular injury pathways within seconds of kinetic exposure. TA1 approaches should ensure countermeasures have target specificity (e.g., 10-fold higher than off-target binding) and binding that prevents downstream heterogeneous molecular, cognitive, and behavioral effects in response to kinetic injury.

Additionally, countermeasures should prevent initiation of injury pathway(s) within five seconds post-exposure to blast or other energetic threats.

1.2.2. TA2: Develop Spatial and Temporal Delivery

The outcome of TA2 will be the precise and temporally relevant delivery and confirmed bioactivity of TA1 therapeutic payloads at sites of injury. Proposers will demonstrate phenotypic outcomes of cell-type and/or regionally specific targeting of countermeasure(s). Performers may choose to leverage approaches including extravasation mechanisms (e.g., oncology approaches), mechanically or chemically triggered mechanisms (e.g., mechanophores, nanoparticles, liposomes), time-resolved release (e.g., chronic or acute), spatially targeted (e.g., cellular and subcellular localized payload release), and/or self-limiting mechanisms to enable the effective and safe delivery of therapeutic payloads. Additionally, TA2 approaches should achieve access to sites of injury (for example, though not exhaustive; CNS, PNS, otoliths, vasculature, etc.) and demonstrate localized bioactivity within five seconds of injury. Lastly, the technology must also provide and maintain protective efficacy for three days, following a single, self-administered, minimally invasive mechanism.

Lastly, chronically bioavailable novel or repurposed drugs, with specific target engagement, theoretical or empirically defined mechanism of action, temporal and spatial localization (without the support of novel delivery systems), and theoretical or demonstrated safety record will be considered.

Proposals that fail to address both TAs will be considered non-conforming and may be rejected without further review.

Fully integrated teams should be assembled early in the proposal process, and the integration of both scientific and managerial responsibilities should be conveyed in the submission. Integrated teams should describe the organizational structure within the team, complete with a dedicated project manager (separate from the principal investigator) to show distribution of responsibilities, lines of communication, and technical tasks throughout the proposal. Teams should structure themselves to support continuous, active engagement with IV&V, U.S. Government stake holders, and regulatory agencies to ensure a path towards clinical translation and regulatory application approvals such as Investigational New Drug (IND) or Emergency Use Authorization (EUA) as appropriate.

1.3. TECHNICAL APPROACH

TA1: Identify, Optimize, and Validate Prophylactic Countermeasures TA1 objectives must achieve countermeasures that prevent the initiation of injury cascades within the first few milliseconds to five seconds of sustaining a kinetic injury.

In TA1, proposed approaches must include utilization of high-resolution molecular structures of targets to inform the design of novel countermeasures. High-resolution structures of countermeasures must demonstrate selective engagements with the target in a manner that preferentially prevents the activation of TBI-associated downstream signaling pathway(s) and subsequent cognitive or behavioral deficits. Countermeasures should show a greater than 10-fold selectivity for the target receptor over closely related anti-targets. TA1 approaches should develop a pipeline with the capability to iteratively optimize the design and activity of countermeasures. Such a pipeline may contain the following components: (1) Informed selection of targets and/or subtype(s) based on evidence for functional roles in TBI; (2) Structural determination of countermeasures and establishment of docking poses conferring signaling bias;

and (3) In vitro or ex vivo assessment of countermeasure-mediated function and validation of predicted signaling bias. For approaches inclusive of drug discovery, proposals should include, at a minimum: state-of-the-art drug discovery approaches, such as virtual docking, predictive toxicity screens, high throughput, and microphysiological assessments in order to identify molecules that prevent targets from triggering downstream signaling injury responses.

Proposers must provide an explicit rationale for the selected target(s), with regard to the involvement of the energetically responsive molecule(s) in the initiation of the TBI. Proposed efforts should include both practical methods for target structure determination, such as X-ray crystallography and/or cryo-electron microscopy, as well as computational approaches, such as homology modeling and molecular dynamics simulations.

Because successful countermeasure development for this program requires reliable discrimination of binding between potentially closely related targets, such as receptors and ion channels, it is critical that the effort employ empirical and iterative determination of target interactions at the atomic scale.

For example, a novel chemotype of predicted value as a lead compound based on activity profiles in vitro must have a clear description of the structural basis of the target interaction determined. In order to prioritize translatability for clinical TBI, target(s) structure determination must begin with human protein sequences and, if derived from animal protein sequence information, deviate minimally from human protein structure.

Proposers developing novel drugs should leverage novel and canonical drug-discovery approaches to design new chemotypes according to predicted structural conformations with associated binding of specific small molecules (at least 10,000 chemotypes docked in silico).

Specifically excluded are methods that rely solely on high-throughput phenotypic screening of existing compound libraries as a first step.

Following the generation of countermeasures from the structure-guided design effort, prioritized countermeasures should be characterized in vitro. The following includes a minimum of SOA techniques that should be included for, but are not limited to, in vitro countermeasure characterization; ligand binding parameters (e.g., Kd, residence time, Bmax, etc.), endogenous ligand displacement, target and or target subtype specificity, profiling the inhibition of target activity during kinetic injury initiated signal transduction (agonist/partial agonist/ antagonist/inverse agonist), and pathway selectivity (e.g., G-protein mediated vs. ß-arrestin).

Proposals should include functional cellular in vitro assays that utilize, at a minimum, appropriate targets, target subtypes, and signal transduction machinery expressed either endogenously by, or introduced into, a relevant model system. Additionally, cellular and multicellular systems should be minimally manipulated and appropriate for the output of the assay. Where appropriate, functional consequences of heterotypic target oligomerization should be explored. For instance, where targets are known to form functional complexes with another receptor or molecule, a model system should be employed that enables formation of the endogenously active complex.

The critical end product of TA1 is a countermeasure with specificity and rapid prophylactic activity following kinetic insult. As such, all TA1-generated countermeasures satisfying the criteria must be evaluated in animal behavioral and cognitive TBI models to establish their utility as prophylactic countermeasures. In addition, countermeasures prioritized from TA1 must have clear mechanisms of action (with demonstrations of causality) described in terms of molecular, physiological, and behavioral phenotypes.

To meet the end goals of the Cornerstone program, TA1 countermeasures should exhibit rapid bioactivity (less than five seconds after kinetic injury) and a normalized effect size exceeding the mean of archetypal compounds studied in human clinical populations (Cohen’s d of 1.4 relative to control). Proposers must also determine the duration (should it exceed three days) of therapeutic efficacy following a single administration of countermeasure.

TA2: Develop Spatial and Temporal Delivery We assert that TBI is driven by injury sustained in specific cell types and/or nervous system regions initiated by energetically responsive receptors. Thus, DARPA is interested in methods under TA2 that investigate the precise spatial and temporal bioeffects of countermeasures in specific cell types, brain regions, or other anatomical structures within the body. Proposals must provide real-time and/or highly resolved approaches for determining the countermeasure delivery to the relevant cell types and regions (e.g., vascular, nervous system (central or peripheral)) along with empirical and/or theoretical rationale for delivery system design.

All proposals must present a plan to demonstrate phenotypic outcomes of cell-type and/or brain-region selective targeting of novel countermeasure(s), compared to non-selective delivery of the same countermeasure(s).

Should proposers include externally triggered delivery approaches, triggers may include mechanical (i.e., shear-stress/strain from the blast/impact event itself) or biochemical stimuli (e.g., pH, calcium, ROS, etc.). Proposed triggered strategies (e.g., mechanical, chemical, etc.)

should detail experimental approaches for demonstrating the precisely timed/localized payload release, while explicitly protecting against inappropriate release (i.e., delivery system leakiness or false-trigger). Energetic distribution and mechanical loads/stresses/strains should be described at a resolution of less than 50 µm3 through finite element modeling, analytical equations, or similar estimation tools.

Finally, the integrated output of TA1 and TA2 (Integrated Countermeasure) will be used to inform the submission of an application for consideration as an IND to the FDA by program end.

In order to do such, proposers must also consider and outline methods to optimize and characterize the formulation of lead compound(s) to satisfy requirements towards an IND to include multiple pre-submissions with the FDA before program completion.

Additional Considerations In light of the heterogeneity in clinical presentations of TBI (molecular, behavioral, cognitive) proposals must include the assessment of therapeutic efficacy (and countermeasure safety) in a minimum of four (4) behavioral and cognitive assessments, and two (2) genetically divergent animal models, as outlined below (See Tests in Section 1.4).

Proposers must demonstrate Integrated Countermeasures are devoid of unintended side effects, such as cognitive or behavioral impairment or stimulation when administered at prophylactic doses in the absence of kinetic injury, no less than 48 hours subsequent to target interactions.

Proposals should also include behavioral and cognitive assessments for repeat dosing regimens.

In addition, where selected targets and or subtype(s) differ in amino acid sequence between humans and animal models such as mice, efforts must include transgenic in vivo model(s) expressing humanized target(s) in order to recapitulate the human target-driven effect(s).

Differences in inhibition of target subtypes and downstream signaling pathways can lead to distinct phenotypic effects. The molecular mechanisms underlying these disparate outcomes are often not well-understood but may be critical for optimization of therapeutic efficacy. Proposers must characterize the downstream signaling events no less than six hours subsequent to target interactions using unbiased profiling methodologies (e.g., protein post-translational modifications (PTMs), synaptoproteomics, and/or transcriptional profiling). Additional relevant information on neurobiological effects such as neuronal morphology (e.g., spine analysis, arborization, and/or neuritogenesis) must also be detailed to further inform the mechanisms underlying phenotypic effects.

Safety and Regulatory Cornerstone-developed Integrated Countermeasures must be characterized for a variety of performance metrics beyond protection against blast. Metrics are detailed in Table 2.

The execution of safety and efficacy assessments will be conducted by performers. These assessments should provide an in-depth understanding of host-toxicity and other factors that will affect performance/safety of both the delivery system (should it be distinct from the countermeasure), and the Integrated Countermeasure (combined countermeasure plus delivery system). Safety testing should demonstrate that preclinical models administered Integrated Countermeasures at prophylactic levels, in the absence of kinetic injury, remain within pre-defined ‘normal physiologies.’ The relevant results and analysis accumulated throughout Safety and Regulatory Testing should be used to update the FDA during pre-submission meetings, incorporating FDA feedback in any updates to the testing plan. This entire dataset will then be used as part of the IND or other submissions prior to human testing in accordance with regulatory guidance.

Proposers must include the following safety testing, at minimum:

TA1

o Physiological responses raised by countermeasures (e.g., long-residence exposure), including organ toxicity (e.g., neuro-, hepato-, reno-, cardio-, gastro-toxicity) and immune responses (e.g., immune cell activation, inflammatory response, and cytokine release);

o In silico ADME predictions, in vitro or ex vivo high throughput toxicology studies; and o Molecular and histological analyses of ex vivo, in vitro, and in vivo studies.

TA2

o Spatial and temporal relevance in vitro and in vivo;

o Integrated Countermeasure temporal bioactivity (in vitro, ex vivo, and in vivo);

o Integrated Countermeasure specific spatial- or specific cell-type bioactivity (in vitro, ex vivo, and in vivo); and o As appropriate:

Trigger (open/close) capability, and associated toxicities (with open- and closed-states) at the level of cell, tissue, and organ;

Physiological responses raised by delivery systems, including immune cell activation, inflammatory response, and cytokine release);

Standard toxicology studies; and molecular and histological analyses of tissues.

Testing and Evaluation (T&E) of the Cornerstone System Performers will be required to evaluate in vitro and in vivo blast models in-house throughout the period of performance. Performer evaluations should include clear molecular, histological, cognitive, and behavioral confirmation of TBI, in addition to clear positive and negative controls for all experimental approaches. Performers are encouraged to compare Countermeasures and Integrated Countermeasures against SOA as appropriate.

IV&V teams will support Cornerstone as a Government-furnished service to independently validate and verify in vitro and in vivo assessments in line with program metrics/milestones. For in vivo test challenges performed by IV&V teams, IV&V teams will define clinically relevant injury models, and behavioral/cognitive assessments following injury.

1.4. PROGRAM SCHEDULE, MILESTONES & METRICS

Table 1: PROGRAM SCHEDULE

Phase I (Base Period) During Phase I, performers will identify and thoroughly characterize a set of first in class countermeasures capable of inhibiting target(s) responsible for initiating injury (molecular, behavioral, cognitive) post-kinetic exposure (e.g., blast) and validate countermeasures in vitro or ex vivo (TA1). Additionally, performers will develop spatial and temporal delivery with in vitro and in vivo-demonstrated specificity (TA2). In both TAs, technologies will be evaluated in vitro and in vivo for safety and toxicity. While tests will focus on select in vivo deliverables, proposals should explicitly outline clear performer-led demonstrations and/or timely reporting of deliverables for all metrics and milestones as listed in Table 2. Lastly, in delivery systems inclusive of triggered release, performers will be required to include false pretense exposures. False pretenses are defined as a trigger or injury asymmetrical to kinetic exposure, such as transient inflammation (e.g., LPS, viral, etc.). False pretenses should demonstrate the (1) safety, and (2) specificity of the countermeasure release under ‘false’ exposure considerations.

Test 1 (9 months) – TA1: Demonstration I: Target causality; II: Spatial relevance; and III:

Temporal relevance.

Performers will demonstrate I) the causality; II) spatial relevance; and III) temporal relevance of their selected targets in two genetically divergent blast-appropriate TBI models (molecular, cognitive, behavioral). In vivo assessments should include time-resolved (e.g., real-time) data collection, and single-cell resolution characterizing TBI. Performers can select up to three unique targets to evaluate in vivo.

Experimental setup, resulting data sets, and analysis will be provided to IV&V teams and DARPA program management in a critical design review at 10 months. The critical review of Test 1 and its outcomes may be leveraged to redesign and modify TA 1 and 2 approaches based on IV&V and DARPA input.

Test 2 (18 months) – TA2: Delivery I: Temporal Precision

Performers will demonstrate in-house temporal delivery in vivo. Test 2 will demonstrate countermeasure delivery and/or payload release and bioactivity within five seconds of blast injury. Experimental assessment must include appropriate in vivo models. In vivo assessment must include detailed, time resolved (e.g., real-time) assessments characterizing the pharmacodynamic and pharmacokinetic payload release and/or bioactivity within five seconds of kinetic injury. Performers may choose to include single-cell resolution to incorporate Test 1 and critical design review feedback as appropriate. Performers may choose payloads to be proof of principal (Luciferase, paramagnets, etc.). Performers with triggerable delivery mechanisms will be required to include a minimum of two false pretenses demonstrating delivery condition specificity. Success will be considered > 80% payload bioactivity within five seconds of blast injury.

Test 3 (20 months) – TA2: Delivery II: Temporal and Spatial Precision

Performed by IV&V, no more than three delivery approaches will be provided to IV&V for testing spatial-specific payload delivery within five seconds of kinetic injury. Delivery systems and/or therapeutic payloads will be assessed for presence and/or bioactivity with cell-or region-specific loci as pre-determined in Test 1. Success will be considered > 80% payload release in targeted loci compared to non-selective delivery of the same payload, within five seconds of kinetic injury. A proof of principal (Luciferase, fluorophores, etc.) payload may be used.

Performers will demonstrate single-cell resolution of spatial specific payload delivery within five seconds of kinetic injury. Performers may choose payloads to be proof of principal (Luciferase, paramagnets, etc.). Performers with triggerable delivery mechanisms will be required to include a minimum of two false pretenses conditions demonstrating delivery spatial and temporal specificity.

Regulatory process

During Phase I, proposers are encouraged to work with the FDA and/or other regulatory agencies. In preparation for active engagement (formal) during Phase II, it is strongly encouraged that all testing in house (inclusive of Safety and Regulatory Testing, Table 1, and

Table 2) is rigorous and carried through with the intent to be used as part of the IND or other submissions prior to human testing in accordance with regulatory guidance.

Phase II (Option One) During Phase II, proposers will focus on integrating and optimizing the capabilities of Integrated Countermeasures. Each team may move forward into Phase II with up to three countermeasures, integrated into no more than two delivery approaches, informed by earlier test results, safety outcomes, and preliminary formulation assessments. In house testing should include a minimum of two genetically divergent, blast appropriate, TBI animal models and a minimum of four total behavioral/cognitive assessments per Test as appropriate. By Test 6, proposers will further narrow to one countermeasure and minimally invasive formulations for IV&V assessment.

Additionally, starting at Test 6, performers will be required to include false pretenses in vivo.

Test 4 (30 months) – TA1: Countermeasure Intervention

Performers will demonstrate in house that administered interventions result in a therapeutic reduction of molecular and cellular injury upon blast exposure. Assessment will consist of prophylactic administration of the countermeasure (either by TA2 delivery systems or IV infusion, lumbar puncture, etc.). Following administration, animals will undergo kinetic injury and highly time-resolved data collection characterizing the downstream molecular and cellular responses of blast injury. No less than three downstream pathway responses must be highly time resolved and detailed (e.g., excitotoxicity, inflammatory, etc.) for a minimum of six hours post-administration. Success will be determined by a consistent > 90% reduction of relevant downstream molecular injury signaling across successive time points (e.g., minutes-hours post injury). Additionally, successful countermeasures will have, at a minimum, demonstrated > 90% reduction in histological markers of neuropathology and neuroinflammation relative to blast-exposed control animals. Performers should include vehicle treatments and SOA comparisons (e.g., barrier provisions such as helmets etc.).

Test 5 (36 months) – TA1: Countermeasure Intervention II

Performers will demonstrate countermeasures result in a therapeutic reduction of behavioral and cognitive deficits upon blast exposure. Up to three countermeasures may be assessed in a minimum of two genetically divergent, blast appropriate, TBI animal models. Assessment will consist of prophylactic administration of countermeasures by drug delivery system or invasive approaches (i.e., IV infusion, lumbar puncture). Following administration, animals will undergo blast injury, and time resolved behavioral and cognitive assessments. No less than four total cognitive and behavioral assessments must be detailed (e.g., learning, memory, vestibular, etc.).

All cognitive and behavioral assessments must have baseline, and acute measurements (e.g., before, and immediately after sustaining blast injury). Success will be determined by a consistent > 90% reduction of behavioral and cognitive impairment across successive time points (e.g., minutes-hours-days post injury) relative to blast exposed control animals. Performers should include vehicle treatments, and SOA comparisons (e.g., barrier provisions such as helmets, etc.).

Test 6 (42 months) – Integrated Countermeasure I

IV&V will assess Integrated Countermeasures for temporal and spatial release after a single administration and two-day residence time. One Integrated Countermeasure will be assessed in a minimum of two genetically divergent, blast appropriate, TBI animal models. Assessment will consist of prophylactic administration of the Integrated Countermeasure by minimally invasive (e.g., patch, eye-drop) or invasive (e.g., IV infusion, subcutaneous injection) means. Animals will undergo blast injury no sooner than one day, and no later than two days post administration.

Performers will be required to demonstrate the absence of toxicity as characterized by molecular, behavioral, and cognitive baselines from therapeutic payloads whether they are activated or not activated. In the inactive and activated state (even if activated without blast exposure), it needs to be shown that there is no toxic effect upon activation no sooner than one day, and no later than two days post administration in a minimum of two genetically divergent animal models. Preliminary efficacy and safety data may be gleaned.

Performers will be required to provide time resolved data collection, along with cell-type, and region-resolved selective targeting/bioactivity, compared to non-temporal and non-spatially targeted delivery of the same therapeutic payload for models of blast exposure, and false pretense exposure. Success will be considered > 90% therapeutic bioactivity at the targeted loci within five seconds of blast injury, and no molecular, cognitive, or behavioral anomalies associated with exposure to false pretenses.

Test 7 (46 months) – Integrated Countermeasure II

IV&V will assess Integrated Countermeasures for protection against blast injury after a single administration and three-day residence time. One Integrated Countermeasure may be assessed in a minimum of two genetically divergent, blast appropriate, TBI animal models. Assessment will consist of prophylactic administration of the Integrated Countermeasure by minimally invasive means (i.e., oral, microneedle patch, eye-drop, etc.). Animals will undergo blast injury no sooner than one day, and no later than three days post administration.

IV&V assessments will include no less than one false pretense concurrent with the three-day window of treatment and blast exposure (i.e., administration, false pretense, and blast exposure, within three-days).

Performers will be required to demonstrate no less than four total cognitive and behavioral measurements (e.g., learning, memory, vestibular, etc.). Cognitive and behavioral measures must assess prophylactic use (e.g., absence of kinetic injury), acute (e.g., within minutes of blast exposure), and chronic (hours-days) after kinetic injury. Success will be determined by (1) zero cognitive or behavioral anomalies during prophylactic use, (2) a consistent > 90% reduction of cognitive and behavioral impairment post blast, across successive time points relative to control animals. Performers should include delivery vehicle treatments and SOA comparisons (e.g., barrier provisions such as helmets, etc.).

Phase III (Option Two) Prior to the end of Phase II (month 46), performers will be required to have engaged with the FDA. Performers should have a clear understanding of all necessary IND/EUA application submission requirements, safety and toxicity data requirements, efficacy demonstrations, Integrated Countermeasure formulations, and plans for technology transition. This phase (Phase III, Months 48-60) will focus on collecting any remaining data, responding to regulatory feedback, and submission of regulatory application(s). For example, submission of package to support FDA IND application, including previous safety and efficacy data from Phases I and II, large animal (e.g., non-human primates) pharmacological and toxicological safety data demonstrating the products are reasonably safe for human testing, and documentation demonstrating capability for good manufacturing practice (GMP) of sufficient quantities of product for clinical testing. Pre-EUA activities are not required but may proceed in parallel to the required FDA approval through IND pathways.

Milestones & Metrics In order for the Government to evaluate the effectiveness of Cornerstone technologies in achieving the stated program objectives, proposers should note that the Government hereby promulgates the following program metrics that may serve as the basis for determining whether satisfactory progress is being made to warrant continued funding of the program. Although the following program metrics are specified, proposers should note that the Government has identified these goals with the intention of bounding the scope of effort, while affording the maximum flexibility, creativity, and innovation in proposing solutions to the stated problem.

Quantitative metrics are expected to vary for each proposer-approach and system. Some exemplary milestones and metrics are included below for proposers to consider, but proposers should adjust accordingly for their given countermeasure and system. Final metrics are to be determined at the time of award negotiation and are subject to DARPA approval. Proposers should note that program metrics may serve as the basis for determining whether satisfactory progress is being made to warrant continued funding of the program (i.e., assessment).

In addition to the tests and system demonstration milestones, performers will be required to participate in program review meetings every six months. All performers will attend these Technical Interchange Meetings (TIMs) to brief their latest results and progress toward program goals. The meetings will include Government participation from interagency stake holders and end-users. Government sidebars will be held to provide individual feedback to the performers and to ensure they are developing relevant technologies. Teleconferences will be held with each team at monthly intervals. Site visits will be conducted at the Program Manager’s discretion.

Table 2 is a summary of the milestones and metrics for each team throughout the Cornerstone program.

Table 2: CORNERSTONE PROGRAM MILESTONES AND METRICS

Phase I (Base): Development and Initial Demonstration Month 1: Virtual program/Phase I kickoff.

Month 3: Institutional Animal Care and Use Committee (IACUC) and Animal Care and

Use Review Office (ACURO) approvals.

Month 9: Performers assess (1) target causality, (2) spatial relevance, and (3) temporal relevance in injury models (Test 1).

Month 10: Critical design review of Test 1 experimental setup, resulting data sets, analysis.

Month 12: Demonstrate cell- or region-specific payload presence and/or bioactivity relative to non-targeted approaches in vitro/ex vivo models.

Month 18: Demonstrate temporal specific payload delivery (less than 5 seconds) relative to non-targeted approaches in vivo (Test 2).

Month 20: Demonstrate spatial specific payload delivery (cell- or region-specific) relative to non-targeted approaches in vivo (Test 3).

Month 22: Deliver Tests 2 and 3 reports.

Phase II (Option): Integrated Countermeasure, Optimization and Efficacy Month 25: Phase II Kick-off meeting.

Month 30: Demonstrate diminished pathological molecular and histopathological responses to kinetic injury (90% vs control) in two or more in vivo models (Test 4).

Month 32: Deliver report for Test 4.

Month 36: Demonstrate reduced cognitive and behavioral deficits (90% vs control) in two or more in vivo models of kinetic injury (Test 5).

Month 38: Deliver report for Test 5.

Month 42: Demonstrate temporal and spatial release of Countermeasure in response to injury after two-day residence time (Test 6).

Month 44: Deliver report for Test 5. Deliver final plan for Phase III studies and documentation.

Month 46: Final demonstration of safety and functionality with protection against molecular, cognitive, and behavioral injury in two or more large animal models (>95% vs uninjured prophylactic treated controls). Demonstration includes a single dose three-day residence time with no safety anomalies.

Month 48: Deliver report for End of Phase Demonstration.

Phase III Option: IND Submission

Month 49: Initiate NHP pre-clinical studies. Initiate demonstration of GMP capability.

Month 55: Deliver mid-phase progress report.

Month 59: Submission of package to support FDA IND application, including previous safety and efficacy data from Phases I and II, large animal (e.g., NHP) pharmacological and toxicological safety data demonstrating the products are reasonably safe for human testing, and documentation demonstrating capability for GMP manufacturing of sufficient quantities of product for clinical testing.

Month 60: Deliver Program Final report.

1.5. GENERAL REQUIREMENTS

Teaming Proposers are responsible for assembling a complete team that has technical expertise, capabilities, and facilities to address all objectives of the program. Proposers must address both TAs which should run in parallel. A complete proposer team should, therefore, not only have the ability to meet the technical challenges of each TA and create an integrated countermeasure, but also have the ability to demonstrate protection against blast and non-lethal false pretenses in relevant preclinical models at appropriate containment levels. It is also encouraged that proposer teams include members that have industrial and commercial experience to aid in focusing on technology research and development strategy for eventual clinical translation. This could include, for example, expertise in medical product development and Good Laboratory Practice (GLP) or GMP manufacturing of medical countermeasures for use in preclinical and clinical settings to effectively navigate the preparatory process for IND/EUA, or equivalent, submission during the program effort. Describe any formal teaming agreements that are required to execute this program. All teams are encouraged to identify a Project Manager to serve as the primary point of contact to communicate with the DARPA Program Manager and Contracting Officer Representative, coordinate effort across performer teams, organize regular performer meetings or discussions, facilitate data sharing, and ensure timely completion of milestones and deliverables.

For teams that are not physically co-located, proposers must articulate how logistical challenges will be overcome to ensure smooth collaboration and an integrated work product.

Animal Subject Research It is anticipated that Animal Subjects Research (ASR) will occur under this effort. As such, proposals should address experience authoring and executing successful ASR protocols.

Furthermore, full proposals must include detailed plans for how the performer intends to acquire all required approvals in a timely manner to meet specified milestones, including local Institutional Animal Care and Use Committee (IACUC) and DoD Animal Care and Use Review Office (ACURO) approvals. Proposers are requested to include a draft IACUC protocol package which will not count toward page limits. Proposers are requested to separate ASR tasks from those that do not require animal subjects research within their Statements of Work.

Ethical, Legal, and Societal Implications (ELSI) DARPA maintains its commitment to ensuring that efforts funded under this BAA adhere to ethical and legal regulations currently in place for Federal and DoD-funded research. Program developments will be discussed with a panel of expert external advisors with expertise in bioethical issues that may emerge as a consequence of advances in biomedical science and technology, including human gene modulation. Proposers to this BAA should address potential ethical, legal, and societal implications of the proposed technology, with a special emphasis on strategies to enable safe, transient, non-permanent Cornerstone technologies.

Deliverables All products, material and otherwise, that will be provided to the government as outcomes from conducted research should be defined as part of the proposal. Performers need to reserve time and budget to fulfill obligations for travel to review meetings and the transmission of report documentation. In addition to the milestones and metrics listed above:

End of Phase reports: At the end of Phase 1 and Phase 2, prior to the initiation of the subsequent phase performers must draft and present to DARPA a written report of all research activities and metrics satisfied. This report should contain as much supporting data as reasonably conveyed.

Monthly financial reports: Performers are required to provide financial status updates.

These reports should be in the form of an editable MS Excel file, and should provide financial data including, but not limited to, the following: program spend plan by phase and task, incurred program expenditures to date by phase and task, and invoiced program expenditures to date by phase and task. The prime performer is to include information for itself and all subawardees/subcontractors.

Monthly technical progress reports: Each month (or as close to as scheduling permits), performers are required to provide research updates. These reports should be in the form of a standardized slide presentation given to DARPA and discussed with the program management team via teleconference. Length and detail level should be at the discretion of the Program Manager.

Quarterly technical reports: Reports…